Mixtures and compositions comprising Paenibacillus strains or metabolites thereof and other biopesticides

ABSTRACT

The present invention relates to novel mixtures comprising, as active components, at least one isolated bacterial strain, which is a member of the genus  Paenibacillus , or a cell-free extract thereof or at least one metabolite thereof, and at least one other biopesticide. The present invention also relates to compositions comprising at least one of such bacterial strains, whole culture broth or a cell-free extract or a fraction thereof or at least one metabolite thereof, and at least one other biopesticide. The present invention also relates to a method of controlling or suppressing plant pathogens or preventing plant pathogen infections by applying such composition. The present invention also relates to mixtures of fusaricidins which are pesticidal metabolites produced by the abovementioned strains, and other biopesticides.

This application is a National Stage application of InternationalApplication No. PCT/EP2017/052535, filed Feb. 6, 2017. This applicationalso claims priority under 35 U.S.C. § 119 to European PatentApplication No. 16154830.0, filed Feb. 9, 2016.

REFERENCE TO SEQUENCE LISTING SUBMITTED VIA EFS-WEB

This application was filed electronically via EFS-Web and includes anelectronically submitted sequence listing in .txt format. The .txt filecontains a sequence listing entitled“13779-1442_2018-12-21_Sequence_Listing” created on Oct. 16, 2018, andis 39,350 bytes in size. The sequence listing contained in this .txtfile is part of the specification and is hereby incorporated byreference herein in its entirety.

FIELD OF THE INVENTION

The present invention relates to novel mixtures comprising, as activecomponents, at least one isolated bacterial strain, which is a member ofthe genus Paenibacillus, or a cell-free extract thereof or at least onemetabolite thereof, and at least one other other biopesticide. Thepresent invention also relates to compositions comprising at least oneof such Paenibacillus strains, whole culture broth or a cell-freeextract or a fraction thereof or at least one metabolite thereof, and atleast one other biopesticide. The present invention also relates to amethod of controlling or suppressing plant pathogens or preventing plantpathogen infections by applying such composition. The present inventionalso relates to mixtures of fusaricidins which are pesticidalmetabolites produced by the abovementioned strains, and otherbiopesticides.

BACKGROUND OF THE INVENTION

In the technical field of controlling phytopathogenic fungi affectingplants or crops it is well known to apply biopesticides, for exampleselected from bacteria, like spore-forming bacteria, or fungi which arenot detrimental to the plant or crop to be treated and which biologicalcontrol agents may be further combined with classical organic chemicalantagonists of plant pathogens.

Biopesticides have been defined as a form of pesticides based onmicro-organisms (bacteria, fungi, viruses, nematodes, etc.) or naturalproducts (compounds or extracts from biological sources) (U.S.Environmental Protection Agency:http://www.epa.gov/pesticides/biopesticides/).

Biopesticides are typically created by growing and concentratingnaturally occurring organisms and/or their metabolites includingbacteria and other microbes, fungi, viruses, nematodes, proteins, etc.They are often considered to be important components of integrated pestmanagement (IPM) programmes, and have received much practical attentionas substitutes to synthetic chemical plant protection products (PPPs).

Biopesticides fall into two major classes, microbial and biochemicalpesticides:

(1) Microbial pesticides consist of bacteria, fungi or viruses (andoften include the metabolites that bacteria and fungi produce).Entomopathogenic nematodes are also classed as microbial pesticides,even though they are multi-cellular.

(2) Biochemical pesticides are naturally occurring substances thatcontrol pests or provide other crop protection uses as defined below,but are relatively non-toxic to mammals.

For controlling phytopathogenic fungi several microbial pesticidescomprising spore-forming bacteria such as Bacillus subtilis have beendescribed earlier, see e. g. WO 1998/050422; WO 2000/029426; WO1998/50422 and WO 2000/58442.

WO 2009/0126473 discloses agriculturally acceptable aqueous compositionscomprising bacterial or fungal spores contained in an aqueous/organicsolvent and which may further comprise insect control agents,pesticides, fungicides or combinations thereof. Spores of bactebacteriaof the genus Bacillus are a preferred species.

WO 2006/017361 discloses compositions for controlling plant pathogensand comprising at least one beneficial bacterium, at least onebeneficial fungus, at least on nutrient and at least one compound whichextends the effective lifetime of such a composition. The group ofbeneficial bacteria e.a. comprises bacteria of Paenibacillus polymyxaand Paenibacillus durum.

WO 1999/059412 discloses a Paenibacillus polymyxa strain PKB1 (bearingATCC accession no. 202127) active against several phytopathogenic fungi.

WO 2011/069227 discloses a P. polymyxa strain JB05-01-1 (bearing ATCCaccession no. PTA-10436) having a highly inhibitory effect againstpathogenic bacteria, pre-dominantly food-borne human pathogenicbacteria.

Raza et al. (Brazilian Arch. Biol. Techol. 53, 1145-1154, 2010; Eur. J.Plant Pathol.125: 471-483, 2009) described a fusaricidin-producingPaenibacillus polymyxa strain SQR-21 effective against Fusariumoxysporum.

Another Paenibacillus polymyxa strain called AC-1 is known fromMicrobial Research 2016 (in press doi:10.1016/j.micres.2016.01.004) andproduct Topseed from Green Biotech Co., Ltd. 45-70 Yadang-ri, Gyoha-EupPaju Kyungki-Do, Korea (South) 413-830.

A further Paenibacillus polymyxa strain presumably called HY96-2 isknown from Biocontrol Science and Technology 24 (4), 426-435 (2014) andshall be marketed under the name KangDiLeiDe by Shanghai Zeyuan MarineBiotechnology Co.,Ltd.

The genome of several Paenibacillus polymyxa strains has been publishedso far: inter alia for strain M-1 (NCBI acc. no. NC_017542; J.Bacteriol. 193 (29), 5862-63, 2011; BMC Microbiol. 13, 137, 2013),strain CR1 (GenBank acc. no. CP006941; Genome Announcements 2 (1), 1,2014) and strain SC2 (GenBank acc. nos. CP002213 and CP002214; NCBI acc.no. NC_014622; J. Bacteriol. 193 (1), 311-312, 2011), for furtherstrains see legend of FIG. 12 herein. The P. polymyxa strain M-1 hasbeen deposited in China General Microbiological Culture CollectionCenter (CGMCC) under acc. no. CGMCC 7581.

In the PCT application PCT/EP2015/067925 (WO 2016/020371) new strains ofthe genus Paenibacillus have been characterized. Said bacterial strainsLu16774, Lu17007 and Lu17015 had been isolated from crop acreage inGermany and deposited under the Budapest Treaty with the DeutscheSammlung von Mikroorganismen und Zellkulturen (DSMZ) on Feb. 20, 2013 byBASF SE, Germany:

-   1) Paenibacillus strain Lu16774 deposited under Accession No. DSM    26969,-   2) Paenibacillus strain Lu17007 deposited under Accession No. DSM    26970, and-   3) Paenibacillus strain Lu17015 deposited under Accession No. DSM    26971.

As used herein, the term Paenibacillus strain is identical to the termPaenibacillus sp. strain and means a bacterial strain form the genusPaenibacillus. The genus Paenibacillus includes all speciesPaenibacillus spp.

In abovementioned PCT/EP2015/067925 (WO 2016/020371), the strainsLu16774, Lu17007 and Lu17015 were determined to belong to the genusPaenibacillus on the following morpholog-morphological and physiologicalobservations (see Example 2.3 in PCT/EP2015/067925 (WO 2016/020371) andherein):

-   -   rod-shaped cells    -   ellipsoidal spores    -   swollen sporangium    -   anaerobic growth    -   fermentation of a variety of sugars including glucose,        arabinose, xylose, mannit, fructose, raffinose, trehalose and        glycerol with acid formation    -   gas production from glucose    -   arginine dihydrolase negative    -   no utilization of citrate    -   no growth in presence of 5% or more sodium chloride    -   production of extracellular hydrolytic enzymes degrading starch,        gelatine, casein and esculin.

Further, these strains Lu16774, Lu17007 and Lu17015 were also determinedto belong to the genus Paenibacillus by 16S rDNA analysis by having thePaenibacillus-specific 22-base sequence in 16S rDNA (5′ to 3′):

5′-TCGATACCCTTGGTGCCGAAGT-3′(see SEQ ID NO:1 (nucleotides 840-861), SEQ ID NO:2 (840-861), SEQ IDNO:3 (844-865) and SEQ ID NO:4 (840-861) in sequence listings ofPCT/EP2015/067925 (WO 2016/020371) and herein).

Further, sequencing of the complete 16S rDNA in comparison to 24different Paenibacillus strains resulted in clustering of the strainsLu16774, Lu17007 and Lu17015 with the type strains of Paenibacillusbrasiliensis, P. kribbensis, P. jamilae, P. peoriae, and P. polymyxa,more preferably to P. peoriae, in particular Paenibacillus peoriaestrain BD-62 (see FIGS. 1 and 2 in PCT/EP2015/067925 (WO 2016/020371)and herein). It is known that P. polymyxa and P. peoriae have 16S rDNAsequence identity values of 99.6 to 99.7% (J. Gen. Appl. Microbiol. 48,281-285 (2002)).

“Percent Identity” or “percent similarity” between two nucleotidesequences means percent identity of the residues over the completelength of the aligned sequences and is determined by comparing twooptimally locally aligned sequences over a comparison window defined bythe length of the local alignment between the two sequences, such as,for example, the identity calculated (for rather similar sequences)after manual alignment with the aid of the program AE2 (Alignment Editor2). Local alignment between two sequences only includes segments of eachsequence that are deemed to be sufficiently similar according to thecriterion that depends on the algorithm used to perform the alignment(e. g. AE2, BLAST, secondary structure of the rRNA molecule or alike).The percentage identity is calculated by determining the number ofpositions at which the identical nucleic acid occurs in both sequencesto yield the number of matched positions, dividing the number of matchedpositions by the total number of positions in the window of comparisonand multiplying the result by 100.

To determine the percent sequence identity of two nucleic acid sequences(e. g. one of the nucleotide sequences of Table 1 and a homologthereof), the sequences are aligned for optimal comparison purposes (e.g. gaps can be introduced in the sequence of one nucleic acid foroptimal alignment with the other nucleic acid). The bases atcorresponding positions are then compared. When a position in onesequence is occupied by the base as the corresponding position in theother sequence then the molecules are identical at that position. It isto be understood that for the purposes of determining sequence identitywhen comparing a DNA sequence to an RNA sequence, a thymidine nucleotideis equivalent to a uracil nucleotide. For alignment, the sequence datawas put into the program AE2(http://iubio.bio.indiana.edu/soft/molbio/unix/ae2.readme), alignedmanually according to the secondary structure of the resulting rRNAmolecule and compared with representative 16S rRNA gene sequences oforganisms belonging to the Firmicutes (Nucl. Acids Res. 27, 171-173,1999). To obtain % identity values for multiple sequences, all sequencesof were aligned with each other (multiple sequence alignment). Further,to obtain % identity values between two sequences over a longer stretchof aligned sequences in comparison to multiple alignment, a manualpairwise sequence alignment was done as described above using AE2(pairwise sequence alignment).

Standardized, automated ribotyping has been performed using the QualiconRiboPrintersystem with the Paenibacillus strains Lu16774, Lu17007 andLu17015 in comparison to the P. peoriae BD-62 using the restrictionenzyme EcoRI resulted in similarity of all three strains Lu16774,Lu17007 and Lu17015 to P. peoriae BD-62 of between 0.24 and 0.5 (seeExample 2.2, FIG. 12 in PCT/EP2015/067925 (WO 2016/020371) and herein).The Paenibacillus strains Lu16774 and Lu17007 were found to belong tothe species Paenibacillus polymyxa.

According to the results of the phylogenetic analysis presented inabovementioned PCT/EP2015/067925 (WO 2016/020371) (FIGS. 12 to 22therein and herein) and unpublished results of Professor Borriss,Germany, the species Paenibacillus polymyxa required a new taxonomicclassification into two subspecies: 1) Paenibacillus polymyxa ssp.polymyxa and 2) Paenibacillus polymyxa ssp. plantarum; and 3) a novelspecies Paenibacillus nov. spec. epiphyticus.

The type strain P. polymyxa DSM 36 together with the P. polymyxa strainsSQR-21, CF05, CICC 10580, NRRL B-30509 and A18 formed in each of themaximum likielihood dendrograms analysed for five conserved housekeeping genes (dnaN, gyrB, recA, recNand rpoA) a separate cluster (FIGS.17-21 of PCT/EP2015/067925 (WO 2016/020371) and herein).

Very similar results have been obtained by determination of the AverageAmino acid Identity (AAI) which is frequently used for determination ofphylogenetic relationship amongst bacterial species. This method isbased on the calculation of the average identity of a core genome onamino acid level (Proc. Natl. Acad. USA 102, 2567-2572, 2005). Accordingto the resulting AAI-matrix in FIG. 22 in PCT/EP2015/067925 (WO2016/020371) and herein, P. polymyxa DSM 36 forms together with the P.polymyxa SQR-21 strain a sub cluster that is different from the twoother sub clusters shown therein.

The strains Lu16674 and Lu17007 together with strain P. polymyxa M-1,1-43, SC2 and Sb3-1 form the second sub cluster in each of the maximumlikielihood dendrograms analysed for five conserved house keeping genes(dnaN, gyrB, recA, recNand rpoA) (FIGS. 17-21). According to AAI-matrixin FIG. 22 in PCT/EP2015/067925 (WO 2016/020371) and herein based on theanalysis of the core genome, this second sub cluster is confirmed by itsrepresentative strains Lu16674 and Lu17007 together with the P. polymyxaM-1 and SC2 strains.

The difference between the two sub clusters was found not so significantto justify a new species, but the AAI identiy levels between therepresentatives of both clusters is of about 97.5% justifying theclassification into two separate subspecies

Thus, it was proposed in PCT/EP2015/067925 (WO 2016/020371) to nominatethe first sub cluster according to the type P. polymyxa strain DSM 36TPaenibacillus polymyxa ssp. polymyxa. Besides strain DSM 36, the P.polymyxa strains SQR-21, CF05, CICC 10580, NRRL B-30509 and A18 shallbelong to the subspecies Paenibacillus polymyxa ssp. polymyxa.

Further, it was proposed to nominate the second sub cluster as novelsubspecies Paenibacillus polymyxa ssp. plantarum. Besides the strainsLu16674 and Lu17007, the P. polymyxa strains M-1, 1-43, SC2 and Sb3-1shall belong to Paenibacillus polymyxa ssp. plantarum.

The strain Lu17015 has only 94.9% AAI identity amongst the genes of thecore genome with the type strain Paenibacillus polymyxa DSM36=ATCC 842(FIG. 22 in PCT/EP2015/067925 (WO 2016/020371) and herein). Thus, thestrain Lu17015 could not have been designated to the speciesPaenibacillus polymyxa nor to any other known Paenibacillus species.Similar values are found for the strains E681 (94.7%) and CR2 (94.9%).Amongst each other, these three strains have at least 98.1% identity(AAI). According to the species definition of Konstantinides and Tiedje(Proc Natl. Acad. Sci. USA. 102, 2567-2572, 2005), the strain Lu17015and also the strains E681 and CR2 can be designated to a novel species.Thus, a new species Paenibacillus spec. nov. epiphyticus has beenproposed in PCT/EP2015/067925 (WO 2016/020371). Consequently, the strainLu17015 belongs to Paenibacillus epiphyticus. It was proposed that saidstrain shall be the type strain. Likewise, the dendrograms based on thesequence comparisons of the five house keeping genes (FIGS. 17-21 inPCT/EP2015/067925 (WO 2016/020371) and herein) showed that this clusteris distant from all other P. polymyxa strains. Besides Lu17015, it wasproposed that the P. polymyxa strains E681, CR2 TD94, DSM 365 and WLY78shall belong to Paenibacillus spec. nov. epiphyticus.

Paenibacillus is known to produce many antibiotic metabolites which arelipopeptides e.g.

polymyxins, octapeptins, polypeptins, pelgipeptins and fusaricidins.Fusaricidins are a group of antibiotics isolated from Paenibacillus spp.from the class of cyclic lipodepsipeptides which often share thefollowing structural features: a macrocyclic ring consisting of 6 aminoacid residues, three of which are L-Thr, D-allo-Thr and D-Ala, as wellas the 15-guanidino-3hydroxypentadecanoic acid tail attached to theN-terminal L-Thr residue by an amide bond (ChemMedChem 7, 871-882, 2012;J. Microbiol. Meth. 85, 175-182, 2011, Table 1 herein). These compoundsare cyclized by a lactone bridge between the N-terminal L-Thr hydroxylgroup and the C-terminal D-Ala carbonyl group. The position of the aminoacid residues within the depsipeptide cycle are usually numberedstarting with the abovementioned L-Thr which itself also carries theGHPD chain and ending with the C-terminal D-Ala. Non-limiting examplesof fusaricidins isolated from Paenibacillus are designated LI-F03,LI-F04, LI-F05, LI-F07 and LI-F08 (J. Antibiotics 40(11), 1506-1514,1987; Heterocycles 53(7), 1533-1549, 2000; Peptides 32, 1917-1923, 2011)and fusaricidins A (also called LI-F04a), B (also called LI-F04b), C(also called LI-F03a) and D (also called LI-F03b) (J. Antibiotics 49(2),129-135, 1996; J. Antibiotics 50(3), 220-228, 1997). The amino acidchain of a fusaricidin is not ribosomally generated but is generated bya non-ribosomal peptide synthetase. Structural formulae of knownfusaricidins are shown in Table 1 (Biotechnol Lett. 34, 1327-1334, 2012;FIG. 1 therein). The compounds designated as LI-F03a, LI-F03b up toLI-F08a and LI-F08b and the fusaricidins of formulae I and 1.1 asdescribed herein are also referred to as fusaricidins LI-F03a, LI-F03bup to LI-F08a and LI-F08b due to their structure within the fusaricidinfamily (see e.g. Table 1).

Among isolated fusaricidin antibiotics, fusaricidin A has shown the mostpromising antimicrobial activity against a variety of clinicallyrelevant fungi and gram-positive bacteria such a Staphylococcus aureus(MIC value range: 0.78-3.12 μg/ml) (ChemMedChem 7, 871-882, 2012). Thesynthesis of fusaricidin analogues that contain 12-guanidino-dodecanoicacid (12-GDA) or 12-amino-dodecanoic acid (12-ADA) instead of naturallyoccurring GHPD has been established but the replacement of GHPD by12-ADA resulted in complete loss of the antimicrobial activity while thereplacement of GHPD by 12-GDA retained antimicrobial activity(Tetrahedron Lett. 47, 8587-8590, 2006; ChemMedChem 7, 871-882, 2012).

TABLE 1 Structures of the fusaricidin family. Fusaricidin X² X³ X⁵ A(LI-F04a) D-Val L-Val D-Asn B (LI-F04b) D-Val L-Val D-Gln C (LI-F03a)D-Val L-Tyr D-Asn D (LI-F03b) D-Val L-Tyr D-Gln LI-F05a D-Val L-IleD-Asn LI-F05b D-Val L-Ile D-Gln LI-F06a D-allo-Ile L-Val D-Asn LI-F06bD-allo-Ile L-Val D-Gln LI-F07a D-Val L-Phe D-Asn LI-F07b D-Val L-PheD-Gln LI-F08a D-Ile L-allo-Ile D-Asn LI-F08b D-Ile L-allo-Ile D-Gln 1A*Ile Tyr Asn 1B* Ile Tyr Asn

wherein an arrow defines a single (amide) bond either between thecarbonyl moiety of GHPD and the amino group of L-Thr (L-threonine) orbetween the carbonyl group of one amino acid and the amino group of aneighboring amino acid, wherein the tip of the arrow indicates theattachment to the amino group of said amino acid L-Thr or of saidneighboring amino acid; andwherein the single line without an arrow head defines a single (ester)bond between the carbonyl group of D-Ala (D-alanine) and the hydroxylgroup of L-Thr; and wherein GHPD is 15-guanidino-3-hydroxypentadecanoicacid.*in case of these two fusaricidins 1A and 1B known from the unpublishedPCT application PCT/EP2015/067925 (WO 2016/020371), the stereoconfiguration of the six amino acids of the cyclic peptide has not beenelucidated.

Fusaricidins A, B, C and D are also reported to inhibit plant pathogenicfungi such as Fusarium oxysporum, Aspergillus niger, Aspergillus oryzae,and Penicillum thomii (J. Antibiotics 49(2), 129-135, 1996; J.Antibiotics 50(3), 220-228, 1997). Fusaricidins such as Li-F05, LI-F07and LI-F08 have been found to have certain antifungal activity againstvarious plant pathogenic fungi such as Fusarium monlliforme, F.oxysporum, F. roseum, Giberella fujkuroi, Helminthosporium sesamum andPenicillium expansum (J. Antibiotics 40(11), 1506-1514, 1987).Fusaricidins also have antibacterial activity to Gram-positive bacteriaincluding Staphylococcus aureus (J. Antibiotics 49, 129-135, 1996; J.Antibiotics 50, 220-228, 1997). In addition, fusaricidins haveantifungal activity against Leptosphaeria maculans which causes blackroot rot of canola (Can. J. Microbiol. 48, 159-169, 2002). Moreover,fusaricidins A and B and two related compounds thereof, whereinD-allo-Thr is bound via its hydroxyl group to an additional alanineusing an ester bridge, produced by certain Paenibacillus strains werefound to induce resistance reactions in cultured parsley cells and toinhibit growth of Fusarium oxysporum (WO 2006/016558; EP 1 788 074 A1).

WO 2007/086645 describes the fusaricidin synthetase enzyme and itsencoding gene as isolated from Paemibacillus polymyxa strain E681 whichenzyme is involved in the synthesis of fusaricidins A, B, C, D, LI-F03,LI-F04, LI-F05, LI-F07 and LI-F08.

In abovementioned PCT/EP2015/067925 (WO 2016/020371) it was found thatthe whole culture broth, the culture medium and cell-free extracts ofthe bacterial strains Lu16774, Lu17007 and Lu17015 show inhibitoryactivity inter alia against Alternaria spp., Botrytis cinerea andPhytophthora infestans. Bioactivity guided fractionation of organicextracts of these strains led to the isolation of two novelfusaricidin-type compounds (herein referred to as fusaricidin 1A and1B), the structure of which were elucidated by 1D- and 2D-NMRspectroscopy as well as mass spectrometry:

Biopesticides for use against crop diseases have already establishedthemselves on a variety of crops. For example, biopesticides alreadyplay an important role in controlling downy mildew diseases. Theirbenefits include: a 0-Day Pre-Harvest Interval and the ability to useunder moderate to severe disease pressure.

However, biopesticides under certain conditions can also havedisadvantages, such as high specificity (requiring an exactidentification of the pest/pathogen and the use of multiple products),slow speed of action (thus making them unsuitable if a pest outbreak isan immediate threat to a crop), variable efficacy due to the influencesof various biotic and abiotic factors (since biopesticides are usuallyliving organisms, which bring about pest/pathogen control by multiplyingwithin the target insect pest/pathogen), and resistance development.

Practical agricultural experience has shown that the repeated andexclusive application of an individual active component in the controlof harmful fungi or insects or other pests leads in many cases to arapid selection of those fungus strains or pest isolates which havedeveloped natural or adapted resistance against the active component inquestion. Effective control of these fungi or pests with the activecomponent in question is then no longer possible.

To reduce the risk of the selection of resistant fungus strains orinsect isolates, mixtures of different active components are nowadaysconventionally employed for controlling harmful fungi or insects orother pests. By combining pestidically active compounds and/orbiopesticides having different mechanisms of action, it is possible toensure successful control over a relatively long period of time.

It is an object of the present invention overcome the abovementioneddisadvantages and to provide, with a view to effective resistancemanagement and effective control of phytopathogenic harmful fungi,insects or other pests or to effective plant growth regulation, atapplication rates which are as low as possible, compositions which, at areduced total amount of pesticides applied, have improved activityagainst the harmful fungi or pests or improved plant growth regulatingactivity (synergistic mixtures) and a broadened activity spectrum, inparticular for certain indications.

One typical problem arising in the field of pest control lies in theneed to reduce the dosage rates of the active ingredient in order toreduce or avoid unfavorable environmental or toxicological effectswhilst still allowing effective pest control. In regard to the instantinvention the term pests embrace animal pests, and harmful fungi.

It was therefore an object of the present invention to providepesticidal mixtures which solve the problems of reducing the dosage rateand/or enhancing the spectrum of activity and/or combining knock-downactivity with prolonged control and/or to resistance management and/orpromoting (increasing) the health of plants.

DESCRIPTION OF THE INVENTION

We have accordingly found that this object is achieved by the mixturesand compositions defined herein, comprising at least one bacterialstrain of the genus Paenibacillus (Paenibacillus strain), or a cell-freeextract thereof or at least one metabolite thereof, and a biopesticideas defined herein.

Thus, the present invention relates to mixtures comprising, as activecomponents

1) at least one Paenibacillus strain, the culture medium or a cell-freeextract thereof or at least one metabolite thereof;

and

2) at least one biopesticide II selected from the groups L1) to L5):

-   -   L1) Microbial pesticides with fungicidal, bactericidal,        viricidal and/or plant defense activator activity: Ampelomyces        quisqualis, Aspergillus flavus, Aureobasidium pullulans,        Bacillus altitudinis, B. amyloliquefaciens, B. megaterium, B.        mojavensis, B. mycoides, B. pumilus, B. simplex, B.        solisalsi, B. subtilis, B. subtilis var. amyloliquefaciens,        Candida oleophlla, C. saitoana, Clavibacter michiganensis        (bacteriophages), Coniothyrium minitans, Cryphonectria        parasitica, Cryptococcus albidus, Dilophosphora alopecuri,        Fusarium oxysporum, Clonostachys rosea f. catenulate (also named        Gliocladium catenulatum), Gliocladium roseum, Lysobacter        antibioticus, L. enzymogenes, Metschnikowia fructicola,        Microdochium dimerum, Microsphaeropsis ochracea, Muscodor albus,        Paenibacillus alvei, Paenibacillus polymyxa, Pantoea vagans,        Penicillium bilaiae, Phlebiopsis gigantea, Pseudomonas sp.,        Pseudomonas chloraphis, Pseudozyma flocculosa, Pichia anomala,        Pythium oligandrum, Sphaerodes mycoparasitica, Streptomyces        griseoviridis, S. lydicus, S. violaceusniger, Talaromyces        flavus, Trichoderma asperelloides, T. asperellum, T.        atroviride, T. fertile, T. gamsii, T. harmatum, T. harzianum, T.        polysporum, T. stromaticum, T. virens, T. viride, Typhula        phacorrhiza, Ulocladium oudemansii, Verticillium dahlia,        zucchini yellow mosaic virus (avirulent strain);    -   L2) Biochemical pesticides with fungicidal, bactericidal,        viricidal and/or plant defense activator activity: harpin        protein, Reynoutria sachalinensis extract;    -   L3) Microbial pesticides with insecticidal, acaricidal,        molluscidal and/or nematicidal activity: Agrobacterium        radiobacter, Bacillus cereus, B. firmus, B. thuringiensis, B.        thuringiensis ssp. aizawai, B. t. ssp. israelensis, B. t. ssp.        galleriae, B. t. ssp. kurstaki, B. t. ssp. tenebrionis,        Beauveria bassiana, B. brongniartii, Burkholderia spp.,        Chromobacterium subtsugae, Cydia pomonella granulovirus (CpGV),        Cryptophlebia leucotreta granulovirus (CrleGV), Flavobacterium        spp., Helicoverpa armigera nucleopolyhedrovirus (HearNPV),        Helicoverpa zea nucleopolyhedrovirus (HzNPV), Helicoverpa zea        single capsid nucleopolyhedrovirus (HzSNPV), Heterorhabditis        bacteriophora, Isaria fumosorosea, Lecaniciffium longisporum, L.        muscarium, Metarhizium anisopliae, Metarhizium ansiopliae var.        anisopliae, M. anisopliae var. acridum, Nomuraea rileyi,        Paecilomyces fumosoroseus, P. lilacinus, Paenibacillus        popilliae, Pasteuria spp., P. nishizawae, P. penetrans, P.        ramosa, P. thornea, P. usgae, Pseudomonas fluorescens,        Spodoptera littoralis nucleopolyhedrovirus (SpliNPV),        Steinernema carpocapsae, S. feltiae, S. kraussei, Streptomyces        galbus, S. microflavus,    -   L4) Biochemical pesticides with insecticidal, acaricidal,        molluscidal, pheromone and/or nematicidal activity: L-carvone,        citral, (E,Z)-7,9-dodecadien-1-yl acetate, ethyl formate,        (E,Z)-2,4-ethyl decadienoate (pear ester),        (Z,Z,E)-7,11,13-hexadecatrienal, heptyl butyrate, isopropyl        myristate, lavanulyl senecioate, cis-jasmone, 2-methyl        1-butanol, methyl eugenol, cis-jasmone, methyl jasmonate,        (E,Z)-2,13-octadecadien-1-ol, (E,Z)-2,13-octadecadien-1-ol        acetate, (E,Z)-3,13-octadecadien-1-ol, R-1-octen-3-ol,        pentatermanone, (E,Z,Z)-3,8,11-tetradecatrienyl acetate,        (Z,E)-9,12-tetradecadien-1-yl acetate, Z-7-tetradecen-2-one,        Z-9-tetradecen-1-yl acetate, Z-11-tetradecenal,        Z-11-tetradecen-1-ol, extract of Chenopodium ambrosiodes, Neem        oil, Quillay extract;    -   L5) Microbial pesticides with plant stress reducing, plant        growth regulator, plant growth promoting and/or yield enhancing        activity: Azospirillum amazonense, A. brasllense, A.        lipoferum, A. irakense, A. halopraeferens, Bradyrhizobium        spp., B. elkanil, B. japonicum, B. liaoningense, B. lupini,        Delfiia acidovorans, Glomus intraradices, Mesorhizobium spp.,        Rhizobium leguminosarum bv. phaseoli, R. I. bb. trifolii, R. I.        bv. viciae, R. tropici, and Sinorhizobium meliloti.

In each of these mixtures the biopesticide II of component 2) isdifferent from the Paenibacillus strain of component 1) chosen.

It is preferred that the mixtures comprise biopesticides II selectedfrom the groups L1) and L2).

According to another embodiment of the invention, mixtures comprisebiopesticides II selected from the groups L3) and L4).

According to another embodiment of the invention, mixtures comprisebiopesticides II selected from the group L5).

According to another embodiment of the invention, mixtures comprisebiopesticides II at least one microbial pesticide selected from thegroups L1) and L5).

The biopesticides from group L1) and/or L2) may also have insecticidal,acaricidal, molluscidal, pheromone, nematicidal, plant stress reducing,plant growth regulator, plant growth promoting and/or yield enhancingactivity. The biopesticides from group L3) and/or L4) may also havefungicidal, bactericidal, viricidal, plant defense activator, plantstress reducing, plant growth regulator, plant growth promoting and/oryield enhancing activity. The biopesticides from group L5) may also havefungicidal, bactericidal, viricidal, plant defense activator,insecticidal, acaricidal, molluscidal, pheromone and/or nematicidalactivity.

Many of these biopesticides have been deposited under deposition numbersmentioned herein (the prefices such as ATCC or DSM refer to the acronymof the respective culture collection, for details see e. g. here:http://www.wfcc.info/ccinfo/collection/by_acronynn/), are referred to inliterature, registered and/or are commercially available: mixtures ofAureobasidium pullulans DSM 14940 and DSM 14941 isolated in 1989 inKonstanz, Germany (e. g. blastospores in Blossom Protect® from bio-fermGmbH, Austria), Azospirillum brasilenase Sp245 originally isolated inwheat reagion of South Brazil (Passo Fundo) at least prior to 1980 (BR11005; e. g. GELFIX® Grannineas from BASF Agricultural Specialties Ltd.,Brazil), A. brasilense strains Ab-V5 and Ab-V6 (e. g. in AzoMax fromNovozymes BioAg Produtos papra Agricultura Ltda., Quattro Barras, Brazilor Sinnbiose-Maiz® from Simbiose-Agro, Brazil; Plant Soil 331, 413-425,2010), Bacillus amyloliquefaciens strain AP-188 (NRRL B-50615 andB-50331; U.S. Pat. No. 8,445,255); B. amyloliquefaciens spp. plantarumD747 isolated from air in Kikugawa-shi, Japan (US 20130236522 A1; FERMBP-8234; e. g. Double Nickel™ 55 WDG from Certis LLC, USA), B.amyloliquefaciens spp. plantarum FZB24 isolated from soil inBrandenburg, Germany (also called SB3615; DSM 96-2; J. Plant Dis. Prot.105, 181-197, 1998; e. g. Taegro® from Novozyme Biologicals, Inc., USA),B. amyloliquefaciens ssp. plantarum FZB42 isolated from soil inBrandenburg, Germany (DSM 23117; J. Plant Dis. Prot. 105, 181-197, 1998;e. g. RhizoVital® 42 from AbiTEP GmbH, Germany), B. amyloliquefaciensssp. plantarum MB1600 isolated from faba bean in Sutton Bonington,Nottinghamshire, U.K. at least before 1988 (also called 1430; NRRLB-50595; US 2012/0149571 A1; e. g. Integral® from BASF Corp., USA), B.amyloliquefaciens spp. plantarum QST-713 isolated from peach orchard in1995 in California, U.S.A. (NRRL B-21661; e. g. Serenade® MAX from BayerCrop Science LP, USA), B. amyloliquefaciens spp. plantarum TJ1000isolated in 1992 in South Dakoda, U.S.A. (also called 1BE; ATCC BAA-390;CA 2471555 A1; e. g. QuickRoots™ from TJ Technologies, Watertown, S.Dak., USA), B. firmus CNCM 1-1582, a variant of parental strain EIP-N1(CNCM 1-1556) isolated from soil of central plain area of Israel (WO2009/126473, U.S. Pat. No. 6,406,690; e. g. Votivo® from BayerCropScience LP, USA), B. pumilus GHA 180 isolated from apple treerhizosphere in Mexico (IDAC 260707-01; e. g. PRO-MIX® BX from PremierHorticulture, Quebec, Canada), B. pumilus INR-7 otherwise referred to asBU-F22 and BU-F33 isolated at least before 1993 from cucumber infestedby Erwinia tracheiphlla (NRRL B-50185, NRRL B-50153; U.S. Pat. No.8,445,255), B. pumilus KFP9F isolated from the rhizosphere of grasses inSouth Africa at least before 2008 (NRRL B-50754; WO 2014/029697; e. g.BAC-UP or FUSION-P from BASF Agricultural Specialities (Pty) Ltd., SouthAfrica), B. pumilus QST 2808 was isolated from soil collected inPohnpei, Federated States of Micronesia, in 1998 (NRRL B-30087; e. g.Sonata® or Ballad® Plus from Bayer Crop Science LP, USA), B. simplex ABU288 (NRRL B-50304; U.S. Pat. No. 8,445,255), B. subtilis FB17 alsocalled UD 1022 or UD10-22 isolated from red beet roots in North America(ATCC PTA-11857; System. Appl. Microbiol. 27, 372-379, 2004; US2010/0260735; WO 2011/109395); B. thuringiensis ssp. aizawai ABTS-1857isolated from soil taken from a lawn in Ephraim, Wis., U.S.A., in 1987(also called ABG-6346; ATCC SD-1372; e. g. XenTari® from BioFa AG,Münsingen, Germany), B. t. ssp. kurstaki ABTS-351 identical to HD-1isolated in 1967 from diseased Pink Bollworm black larvae inBrownsville, Tex., U.S.A. (ATCC SD-1275; e. g. Dipel® DF from ValentBioSciences, Ill., USA), B. t. ssp. kurstaki SB4 isolated from E.saccharina larval cadavers (NRRL B-50753; e. g. Beta Pro® from BASFAgricultural Specialities (Pty) Ltd., South Africa), B. t. ssp.tenebrionis NB-176-1, a mutant of strain NB-125, a wild type strainisolated in 1982 from a dead pupa of the beetle Tenebrio molitor (DSM5480; EP 585 215 B1; e. g. Novodor® from Valent BioSciences,Switzerland), Beauveria bassiana GHA (ATCC 74250; e. g. BotaniGard®22WGP from Laverlam Int. Corp., USA), B. bassiana JW-1 (ATCC 74040; e.g. Naturalis® from CBC (Europe) S.r.I., Italy), B. bassiana PPRI 5339isolated from the larva of the tortoise beetle Conchyloctenia punctata(NRRL 50757; e. g. Broad Band® from BASF Agricultural Specialities (Pty)Ltd., South Africa), Bradyrhizobium elkanii strains SEMIA 5019 (alsocalled 29W) isolated in Rio de Janeiro, Brazil and SEMIA 587 isolated in1967 in the State of Rio Grande do Sul, from an area previouslyinoculated with a North American isolate, and used in commercialinoculants since 1968 (Appl. Environ. Microbiol. 73(8), 2635, 2007; e.g. GELFIX 5 from BASF Agricultural Specialties Ltd., Brazil), B.japonicum 532c isolated from Wisconsin field in U.S.A. (Nitragin 61A152;Can. J. Plant. Sci. 70, 661-666, 1990; e. g. in Rhizoflo®, Histick®,Hicoat® Super from BASF Agricultural Specialties Ltd., Canada), B.japonicum E-109 variant of strain USDA 138 (INTA E109, SEMIA 5085; Eur.J. Soil Biol. 45, 28-35, 2009; Biol. Fertil. Soils 47, 81-89, 2011); B.japonicum strains deposited at SEMIA known from Appl. Environ.Microbiol. 73(8), 2635, 2007: SEMIA 5079 isolated from soil in Cerradosregion, Brazil by Embrapa-Cerrados used in commercial inoculants since1992 (CPAC 15; e. g. GELFIX 5 or ADHERE 60 from BASF AgriculturalSpecialties Ltd., Brazil), B. japonicum SEMIA 5080 obtained under labcondtions by Embrapa-Cerrados in Brazil and used in commercialinoculants since 1992, being a natural variant of SEMIA 586 (CB1809)originally isolated in U.S.A. (CPAC 7; e. g. GELFIX 5 or ADHERE 60 fromBASF Agricultural Specialties Ltd., Brazil); Burkholderia sp. A396isolated from soil in Nikko, Japan, in 2008 (NRRL B-50319; WO2013/032693; Marrone Bio Innovations, Inc., USA), Coniothyrium minitansCON/M/91-08 isolated from oilseed rape (WO 1996/021358; DSM 9660; e. g.Contans® WG, Intercept® WG from Bayer CropScience AG, Germany), harpin(alpha-beta) protein (Science 257, 85-88, 1992; e. g. Messenger™ orHARP-N-Tek from Plant Health Care plc, U.K.), Helicoverpa armigeranucleopolyhedrovirus (HearNPV) (J. Invertebrate Pathol. 107, 112-126,2011; e. g. Helicovex® from Adermatt Biocontrol, Switzerland; Diplomata®from Koppert, Brazil; Vivus® Max from AgBiTech Pty Ltd., Queensland,Australia), Helicoverpa zea single capsid nucleopolyhedrovirus (HzSNPV)(e. g. Gemstar® from Certis LLC, USA), Helicoverpa zeanucleopolyhedrovirus ABA-NPV-U (e. g. Heligen® from AgBiTech Pty Ltd.,Queensland, Australia), Heterorhabditis bacteriophora (e. g. Nemasys® Gfrom BASF Agricultural Specialities Limited, UK), Isaria fumosoroseaApopka-97 isolated from mealy bug on gynura in Apopka, Fla., U.S.A.(ATCC 20874; Biocontrol Science Technol. 22(7), 747-761, 2012; e. g.PFR-97™ or PreFeRal® from Certis LLC, USA), Metarhizium anisopliae var.anisopliae F52 also called 275 or V275 isolated from codling moth inAustria (DSM 3884, ATCC 90448; e. g. Met52® Novozymes Biologicals BioAgGroup, Canada), Metschnikowia fructicola 277 isolated from grapes in thecentral part of Israel (U.S. Pat. No 6,994,849; NRRL Y-30752; e. g.formerly Shemer® from Agrogreen, Israel), Paecllomyces llacinus 251isolated from infected nematode eggs in the Philippines (AGAL 89/030550;WO1991/02051; Crop Protection 27, 352-361, 2008; e. g. BioAct® fromBayer CropScience AG, Germany and MeloCon® from Certis, USA),Paenibaclllus alvei NAS6G6 isolated from the rhizosphere of grasses inSouth Africa at least before 2008 (WO 2014/029697; NRRL B-50755; e.g.BAC-UP from BASF Agricultural Specialities (Pty) Ltd., South Africa),Pasteuria nishizawae Pn1 isolated from a soybean field in the mid-2000sin Illinois, U.S.A. (ATCC SD-5833; Federal Register 76(22), 5808, Feb.2, 2011; e.g. Clariva™ PN from Syngenta Crop Protection, LLC, USA),Penicillium bilaiae (also called P. bilaii) strains ATCC 18309 (=ATCC74319), ATCC 20851 and/or ATCC 22348 (=ATCC 74318) originally isolatedfrom soil in Alberta, Canada (Fertilizer Res. 39, 97-103, 1994; Can. J.Plant Sci. 78(1), 91-102, 1998; U.S. Pat. No. 5,026,417, WO 1995/017806;e. g. Jump Start®, Provide® from Novozymes Biologicals BioAg Group,Canada), Reynoutria sachalinensis extract (EP 0307510 B1; e. g. Regalia®SC from Marrone BioInnovations, Davis, Calif., USA or Milsana® fromBioFa AG, Germany), Steinernma carpocapsae (e. g. Millenium® from BASFAgricultural Specialities Limited, UK), S. feltiae (e. g. Nemashield®from BioWorks, Inc., USA; Nemasys® from BASF Agricultural SpecialitiesLimited, UK), Streptomyces microflavus NRRL B-50550 (WO 2014/124369;Bayer CropScience, Germany), Trichoderma asperelloides JM41R isolated inSouth Africa (NRRL 50759; also referred to as T. fertile; e. g.Trichoplus® from BASF Agricultural Specialities (Pty) Ltd., SouthAfrica), T. harzianum T-22 also called KRL-AG2 (ATCC 20847; BioControl57, 687-696, 2012; e. g. Plantshield® from BioWorks Inc., USA or SabrEx™from Advanced Biological Marketing Inc., Van Wert, Ohio, USA).

According to one embodiment of the inventive mixtures, the at least onepesticide II is selected from the groups L1) to L5):

-   -   L1) Microbial pesticides with fungicidal, bactericidal,        viricidal and/or plant defense activator activity: Aureobasidium        pullulans DSM 14940 and DSM 14941 (L1.1), Bacillus        amyloliquefaciens AP-188 (L.1.2), B. amyloliquefaciens ssp.        plantarum D747 (L.1.3),B. amyloliquefaciens ssp. plantarum FZB24        (L.1.4), B. amyloliquefaciens ssp. plantarum FZB42 (L.1.5), B.        amyloliquefaciens ssp. plantarum MBI600 (L.1.6), B.        amyloliquefaciens ssp. plantarum QST-713 (L.1.7), B.        amyloliquefaciens ssp. plantarum TJ1000 (L.1.8), B. pumilus GB34        (L.1.9), B. pumilus GHA 180 (L.1.10), B. pumilus INR-7        (L.1.11), B. pumilus KFP9F (L.1.12), B. pumilus QST 2808        (L.1.13), B. simplex ABU 288 (L.1.14), B. subtilis FB17        (L.1.15), Coniothyrium minitans CON/M/91-08 (L.1.16),        Metschnikowia fructicola NRRL Y-30752 (L.1.17), Paenibacillus        alvei NAS6G6 (L.1.18), Penicillium bilaiae ATCC 22348        (L.1.19), P. bilaiae ATCC 20851 (L.1.20), Penicillum bilaiae        ATCC 18309 (L.1.21), Streptomyces microflavus NRRL B-50550        (L.1.22), Trichoderma asperelloides JM41R (L.1.23), T. harzianum        T-22 (L.1.24);    -   L2) Biochemical pesticides with fungicidal, bactericidal,        viricidal and/or plant defense activator activity: harpin        protein (L.2.1), Reynoutria sachalinensis extract (L.2.2);    -   L3) Microbial pesticides with insecticidal, acaricidal,        molluscidal and/or nematicidal activity: Bacillus firmus I-1582        (L.3.1); B. thuringiensis ssp. aizawai ABTS-1857 (L.3.2), B. t.        ssp. kurstaki ABTS-351 (L.3.3), B. t. ssp. kurstaki SB4        (L.3.4), B. t. ssp. tenebrionis NB-176-1 (L.3.5), Beauveria        bassiana GHA (L.3.6), B. bassiana JW-1 (L.3.7), B. bassiana PPRI        5339 (L.3.8), Burkholderia sp. A396 (L.3.9), Helicoverpa        armigera nucleopolyhedrovirus (HearNPV) (L.3.10), Helicoverpa        zea nucleopolyhedrovirus (HzNPV) ABA-NPV-U (L.3.11), Helicoverpa        zea single capsid nucleopolyhedrovirus (HzSNPV) (L.3.12),        Heterohabditis bacteriophora (L.3.13), Isaria fumosorosea        Apopka-97 (L.3.14), Metarhizium anisopliae var. anisopliae F52        (L.3.15), Paecilomyces lilacinus 251 (L.3.16), Pasteuria        nishizawae Pn1 (L.3.17), Steinernema carpocapsae (L.3.18), S.        feltiae (L.3.19);    -   L4) Biochemical pesticides with insecticidal, acaricidal,        molluscidal, pheromone and/or nematicidal activity: cis-jasmone        (L.4.1), methyl jasmonate (L.4.2), Quillay extract (L.4.3);    -   L5) Microbial pesticides with plant stress reducing, plant        growth regulator, plant growth promoting and/or yield enhancing        activity: Azospirillum brasilense Ab-V5 and Ab-V6 (L.5.1), A.        brasilense Sp245 (L.5.2), Bradyrhizobium elkanii SEMIA 587        (L.5.3), B. elkanii SEMIA 5019 (L.5.4), B. japonicum 532c        (L.5.5), B. japonicum E-109 (L.5.6), B. japonicum SEMIA 5079        (L.5.7), B. japonicum SEMIA 5080 (L.5.8).

The present invention furthermore relates to agrochemical compositionscomprising a mixture of component 1) as defined herein and at least onebiopesticide II (component 2), in particular at least one biopesticideselected from the groups L1) and L2), as described above, and if desiredat least one suitable auxiliary.

The present invention furthermore relates to agrochemical compositionscomprising a mixture of of component 1) as defined herein and at leastone biopesticide selected from the group L) (component 2), in particularat least one biopesticide selected from the groups L3) and L4), asdescribed above, and if desired at least one suitable auxiliary.

Preference is also given to mixtures comprising as biopesticide II(component 2) a microbial pesticide selected from the groups L1), L3)and L5), preferably selected from strains denoted above as (L.1.2),(L.1.3), (L.1.4), (L.1.5), (L.1.6), (L.1.7), (L.1.8), (L.1.10),(L.1.11), (L.1.12), (L.1.13), (L.1.14), (L.1.15), (L.1.17), (L.1.18),(L.1.19), (L.1.20), (L.1.21), (L.3.1); (L.3.9), (L.3.16), (L.3.17),(L.5.1), (L.5.2), (L.5.3), (L.5.4), (L.5.5), (L.5.6), (L.5.7), (L.5.8);(L.4.2), and (L.4.1); even more preferably selected from (L.1.2),(L.1.6), (L.1.7), (L.1.8), (L.1.11), (L.1.12), (L.1.13), (L.1.14),(L.1.15), (L.1.18), (L.1.19), (L.1.20), (L.1.21), (L.3.1); (L.3.9),(L.3.16), (L.3.17), (L.5.1), (L.5.2), (L.5.5), (L.5.6); (L.4.2), and(L.4.1). These mixtures are particularly suitable for treatment ofpropagation materials, i. e. seed treatment purposes and likewise forsoil treatment. These seed treatment mixtures are particularly suitablefor crops such as cereals, corn and leguminous plants such as soybean.

Preference is also given to mixtures comprising as pesticide II(component 2) a biopesticide selected from the groups L1), L3) and L5),preferably selected from strains denoted above as (L1.1), (L.1.2),(L.1.3), (L.1.6), (L.1.7), (L.1.9), (L.1.11), (L.1.12), (L.1.13),(L.1.14), (L.1.15), (L.1.17), (L.1.18), (L.1.22), (L.1.23), (L.1.24),(L.2.2); (L.3.2), (L.3.3), (L.3.4), (L.3.5), (L.3.6), (L.3.7), (L.3.8),(L.3.10), (L.3.11), (L.3.12), (L.3.13), (L.3.14), (L.3.15), (L.3.18),(L.3.19); (L.4.2), even more preferably selected from (L.1.2), (L.1.7),(L.1.11), (L.1.13), (L.1.14), (L.1.15), (L.1.18), (L.1.23), (L.3.3),(L.3.4), (L.3.6), (L.3.7), (L.3.8), (L.3.10), (L.3.11), (L.3.12),(L.3.15), and (L.4.2). These mixtures are particularly suitable forfoliar treatment. These mixtures for foliar treatment are particularlysuitable for vegetables, fruits, vines, cereals, corn, leguminous cropssuch as soybeans.

Many of these biopesticides have been deposited under deposition numbersmentioned herein (the prefices such as ATCC or DSM refer to the acronymof the respective culture collection, for details see e. g. here:http://www.wfcc.info/ccinfo/collection/by_acronynn/), are referred to inliterature, registered and/or are commercially available: mixtures ofAureobasidium pullulans DSM 14940 and DSM 14941 isolated in 1989 inKonstanz, Germany (e. g. blastospores in Blossom Protect® from bio-fernnGmbH, Austria), Bacillus amyloliquefaciens strain AP-188 (NRRL B-50615and B-50331; U.S. Pat. No. 8,445,255); B. amyloliquefaciens spp.plantarum D747 isolated from air in Kikugawa-shi, Japan (US 20130236522A1; FERM BP-8234; e. g. Double Nickel™ 55 WDG from Certis LLC, USA), B.amyloliquefaciens spp. plantarum FZB24 isolated from soil inBrandenburg, Germany (also called SB3615; DSM 96-2; J. Plant Dis. Prot.105, 181-197, 1998; e. g. Taegro® from Novozyme Biologicals, Inc., USA),B. amyloliquefaciens ssp. plantarum FZB42 isolated from soil inBrandenburg, Germany (DSM 23117; J. Plant Dis. Prot. 105, 181-197, 1998;e. g. RhizoVital® 42 from AbiTEP GmbH, Germany), B. amyloliquefaciensspp. plantarum MBI600 isolated from faba bean in Sutton Bonington,Nottinghamshire, U.K. at least before 1988 (also called 1430; NRRLB-50595; US 2012/0149571 A1; e. g. Integral® from BASF Corp., USA), B.amyloliquefaciens spp. plantarum QST-713 isolated from peach orchard in1995 in California, U.S.A. (NRRL B-21661; e. g. Serenade® MAX from BayerCrop Science LP, USA), B. amyloliquefaciens spp. plantarum TJ1000isolated in 1992 in South Dakoda, U.S.A. (also called 1BE; ATCC BAA-390;CA 2471555 A1; e. g. QuickRoots™ from TJ Technologies, Watertown, S.Dak., USA), B. firmus CNCM 1-1582, a variant of parental strain EIP-N1(CNCM 1-1556) isolated from soil of central plain area of Israel (WO2009/126473, U.S. Pat. No. 6,406,690; e. g. Votivo® from BayerCropScience LP, USA), B. pumllusGHA 180 isolated from apple treerhizosphere in Mexico (IDAC 260707-01; e. g. PRO-MIX® BX from PremierHorticulture, Quebec, Canada), B. pumllus INR-7 otherwise referred to asBU-F22 and BU-F33 isolated at least before 1993 from cucumber infestedby Erwin/a tracheiphlla(NRRL B-50185, NRRL B-50153; U.S. Pat. No.8,445,255), B. pumllusQST 2808 was isolated from soil collected inPohnpei, Federated States of Micronesia, in 1998 (NRRL B-30087; e. g.Sonata® or Ballad® Plus from Bayer Crop Science LP, USA), B. simplex ABU288 (NRRL B-50304; U.S. Pat. No. 8,445,255), B. subas FB17 also calledUD 1022 or UD10-22 isolated from red beet roots in North America (ATCCPTA-11857; System. Appl. Microbiol. 27, 372-379, 2004; US 2010/0260735;WO 2011/109395); B. thuringiensIS ssp. aizawai ABTS-1857 isolated fromsoil taken from a lawn in Ephraim, Wis., U.S.A., in 1987 (also calledABG-6346; ATCC SD-1372; e. g. XenTari® from BioFa AG, Munsingen,Germany), B. t. ssp. kurstak/ABTS-351 identical to HD-1 isolated in 1967from diseased Pink Bollworm black larvae in Brownsville, Tex., U.S.A.(ATCC SD-1275; e. g. Dipel® DF from Valent BioSciences, Ill., USA), B.t. ssp. tenebrion1S NB-176-1, a mutant of strain NB-125, a wild typestrain isolated in 1982 from a dead pupa of the beetle Tenebrio molitor(DSM 5480; EP 585 215 B1; e. g. Novodor® from Valent BioSciences,Switzerland), Beauver/a bass/ana GHA (ATCC 74250; e. g.

BotaniGard® 22WGP from Laverlam Int. Corp., USA), B. bass/ana JW-1 (ATCC74040; e. g. Naturalis® from CBC (Europe) S.r.I., Italy), Burkho/der/asp. A396 isolated from soil in Nikko, Japan, in 2008 (NRRL B-50319; WO2013/032693; Marrone Bio Innovations, Inc., USA), Coniothyrium minitansCON/M/91-08 isolated from oilseed rape (WO 1996/021358; DSM 9660; e. g.Contans® WG, Intercept® WG from Bayer CropScience AG, Germany), harpin(alpha-beta) protein (Science 257, 85-88, 1992; e. g. Messenger™ orHARP-N-Tek from Plant Health Care plc, U.K.), Helicoverpa armigeranucleopolyhedrovirus (HearNPV) (J. Invertebrate Pathol. 107, 112-126,2011; e. g. Helicovex0 from Adermatt Biocontrol, Switzerland;Diplonnata0 from Koppert, Brazil; Vivus® Max from AgBiTech Pty Ltd.,Queensland, Australia), Helicoverpa zea single capsidnucleopolyhedrovirus (HzSNPV) (e. g. Gemstar® from Certis LLC, USA),

Helicoverpa zea nucleopolyhedrovirus ABA-NPV-U (e. g. Heligen® fromAgBiTech Pty Ltd., Queensland, Australia), Heterorhabd/t/s bacter/ophora(e. g. Nemasys® G from BASF Agricultural Specialities Limited, UK),Isaria fumosorosea Apopka-97 isolated from mealy bug on gynura inApopka, Fla., U.S.A. (ATCC 20874; Biocontrol Science Technol. 22(7),747-761, 2012; e. g. PFR-97™ or PreFeRal® from Certis LLC, USA),Metarhizium amsopliae var. anisopilae F52 also called 275 or V275isolated from codling moth in Austria (DSM 3884, ATCC 90448; e. g.Met52® Novozymes Biologicals BioAg Group, Canada), Metschn/kow/afruct/cola 277 isolated from grapes in the central part of Israel (U.S.Pat. No. 6,994,849; NRRL Y-30752; e. g. formerly Shemer® from Agrogreen,Israel), Paecllomyces ilacinus 251 isolated from infected nematode eggsin the Philippines (AGAL 89/030550; WO1991/02051; Crop Protection 27,352-361, 2008; e. g. BioAct® from Bayer CropScience AG, Germany andMeloCon® from Certis, USA), Pasteur/a n/shizawae Pn1 isolated from asoybean field in the mid-2000s in Illinois, U.S.A. (ATCC SD-5833;Federal Register 76(22), 5808, Feb. 2, 2011; e.g. Clariva™ PN fromSyngenta Crop Protection, LLC, USA), Penicilium bilaiae (also called P.bilaii) strains ATCC 18309 (=ATCC 74319), ATCC 20851 and/or ATCC 22348(=ATCC 74318) originally isolated from soil in Alberta, Canada(Fertilizer Res. 39, 97-103, 1994; Can. J. Plant Sci. 78(1), 91-102,1998; U.S. Pat. No. 5,026,417, WO 1995/017806; e. g. Jump Start®,Provide® from Novozymes Biologicals BioAg Group, Canada), Reynoutriasachalinensis extract (EP 0307510 B1; e. g. Regalia® SC from MarroneBioInnovations, Davis, Calif., USA or Milsana® from BioFa AG, Germany),Steinemema carpocapsae(e. g. Millenium® from BASF AgriculturalSpecialities Limited, UK), S. feltiae (e. g. Nemashield® from BioWorks,Inc., USA; Nemasys® from BASF Agricultural Specialities Limited, UK),Streptomyces microflavus NRRL B-50550 (WO 2014/124369; BayerCropScience, Germany), T harzianurn T-22 also called KRL-AG2 (ATCC20847; BioControl 57, 687-696, 2012; e. g. Plantshield® from BioWorksInc., USA or SabrEx™ from Advanced Biological Marketing Inc., Van Wert,Ohio, USA).

According to one embodiment of the inventive mixtures, the at least onepesticide II is selected from the groups L1) to L4):

-   -   L1) Microbial pesticides with fungicidal, bactericidal,        viricidal and/or plant defense activator activity: Aureobasidium        pullulans DSM 14940 and DSM 14941 (L1.1), Bacillus        amyloliquefaciens AP-188 (L.1.2), B. amyloliquefaciens ssp.        plantarum D747 (L.1.3), B. amyloliquefaciens ssp. plantarum        FZB24 (L.1.4), B. amyloliquefaciens ssp. plantarum FZB42        (L.1.5), B. amyloliquefaciens ssp. plantarum MBI600 (L.1.6), B.        amyloliquefaciens ssp. plantarum QST-713 (L.1.7), B.        amyloliquefaciens ssp. plantarum TJ1000 (L.1.8), B. pumilus GB34        (L.1.9), B. pumilus GHA 180 (L.1.10), B. pumilus INR-7        (L.1.11), B. pumilus QST 2808 (L.1.13), B. simplex ABU 288        (L.1.14), B. subtilis FB17 (L.1.15), Coniothyrium minitans        CON/M/91-08 (L.1.16), Metschnikowia fructicola NRRL Y-30752        (L.1.17), Penicilium bilaiae ATCC 22348 (L.1.19), P. bilaiaeATCC        20851 (L.1.20), Penicillium bilaiae ATCC 18309 (L.1.21),        Streptomyces microflavusNRRL B-50550 (L.1.22), Trichoderma        harzianum T-22 (L.1.24);    -   L2) Biochemical pesticides with fungicidal, bactericidal,        viricidal and/or plant defense activator activity: harpin        protein (L.2.1), Reynoutria sachalinensis extract (L.2.2);    -   L3) Microbial pesticides with insecticidal, acaricidal,        molluscidal and/or nematicidal activity: Bacillus firmus I-1582        (L.3.1); B. thuringiensis ssp. aizawai ABTS-1857 (L.3.2), B. t.        ssp. kurstaki ABTS-351 (L.3.3), B. t. ssp. tenebrionis NB-176-1        (L.3.5), Beauveria bassiana GHA (L.3.6), B. bassiana JW-1        (L.3.7), Burkholderia sp. A396 (L.3.9), Helicoverpa armigera        nucleopolyhedrovirus (HearNPV) (L.3.10), Helicoverpa zea        nucleopolyhedrovirus (HzNPV) ABA-NPV-U (L.3.11), Helicoverpa zea        single capsid nucleopolyhedrovirus (HzSNPV) (L.3.12),        Heterohabditis bacteriophora (L.3.13), Isaria fumosorosea        Apopka-97 (L.3.14), Metarhizium anisopliae var. anisopliae F52        (L.3.15), Paecilomyces lilacinus 251 (L.3.16), Pasteuria        nishizawae Pn1 (L.3.17), Steinernema carpocapsae (L.3.18), S.        feltiae (L.3.19);    -   L4) Biochemical pesticides with insecticidal, acaricidal,        molluscidal, pheromone and/or nematicidal activity: cis-jasmone        (L.4.1), methyl jasmonate (L.4.2), Quillay extract (L.4.3).

The present invention furthermore relates to agrochemical compositionscomprising a mixture of XXX (component 1) and at least one biopesticideselected from the group L) (component 2), in particular at least onebiopesticide selected from the groups L1) and L2), as described above,and if desired at least one suitable auxiliary.

The present invention furthermore relates to agrochemical compositionscomprising a mixture of XXX (component 1) and at least one biopesticideselected from the group L) (component 2), in particular at least onebiopesticide selected from the groups L3) and L4), as described above,and if desired at least one suitable auxiliary.

Preference is also given to mixtures comprising as pesticide II(component 2) a biopesticide selected from the groups L1), L3) and L5),preferably selected from strains denoted above as (L.1.2), (L.1.3),(L.1.4), (L.1.5), (L.1.6), (L.1.7), (L.1.8), (L.1.10), (L.1.11),(L.1.13), (L.1.14), (L.1.15), (L.1.17), (L.1.19), (L.1.20), (L.1.21),(L.3.1); (L.3.9), (L.3.16), (L.3.17), (L.4.2), and (L.4.1); even morepreferably selected from (L.1.2), (L.1.6), (L.1.7), (L.1.8), (L.1.11),(L.1.13), (L.1.14), (L.1.15), (L.1.19), (L.1.20), (L.1.21), (L.3.1);(L.3.9), (L.3.16), (L.3.17); (L.4.2), and (L.4.1). These mixtures areparticularly suitable for treatment of propagation mateials, i. e. seedtreatment purposes and likewise for soil treatment. These seed treatmentmixtures are particularly suitable for crops such as cereals, corn andleguminous plants such as soybean.

Preference is also given to mixtures comprising as pesticide II(component 2) a biopesticide selected from the groups L1), L3) and L5),preferably selected from strains denoted above as (L1.1), (L.1.2),(L.1.3), (L.1.6), (L.1.7), (L.1.9), (L.1.11), (L.1.13), (L.1.14),(L.1.15), (L.1.17), (L.1.22), (L.1.24), (L.2.2); (L.3.2), (L.3.3),(L.3.5), (L.3.6), (L.3.7), (L.3.10), (L.3.11), (L.3.12), (L.3.13),(L.3.14), (L.3.15), (L.3.18), (L.3.19); (L.4.2), even more preferablyselected from (L.1.2), (L.1.7), (L.1.11), (L.1.13), (L.1.14), (L.1.15),(L.3.3), (L.3.6), (L.3.7), (L.3.10), (L.3.11), (L.3.12), (L.3.15), and(L.4.2). These mixtures are particularly suitable for foliar treatment.These mixtures for foliar treatment are particularly suitable forvegetables, fruits, vines, cereals, corn, leguminous crops such assoybeans.

According to one embodiment, the microbial pesticides selected fromgroups L1), L3) and L5) embrace not only the isolated, pure cultures ofthe respective microorganism as defined herein, but also its cell-freeextract, its suspensions in a whole broth culture or as ametabolite-containing culture medium or a purified metabolite obtainedfrom a whole broth culture of the microorganism.

A further embodiment relates to mixtures comprising as component 1) awhole culture broth, a culture medium or a cell-free extract or afraction or at least one metabolite of at least one of themicroorganisms as defined above which preferably exhibit antagonisticactivity against at least one plant pathogen.

As used herein, “whole culture broth” refers to a liquid culture of amicroorganism containing vegetative cells and/or spores suspended in theculture medium and optionally metabolites produced by the respectivemicroorganism.

As used herein, “culture medium”, refers to a medium obtainable byculturing the microorganism in said medium, preferably a liquid broth,and remaining when cells grown in the medium are removed, e. g., thesupernatant remaining when cells grown in a liquid broth are removed bycentrifugation, filtration, sedimentation, or other means well known inthe art; comprising e. g. metabolites produced by the respectivemicroorganism and secreted into the culture medium. The “culture medium”sometimes also referred to as “supernatant” can be obtained e. g. bycentrifugation at temperatures of about 2 to 30° C. (more preferably attemp-temperatures of 4 to 20° C.) for about 10 to 60 min (morepreferably about 15 to 30 min) at about 5,000 to 20,000×g (morepreferably at about 15,000×g).

As used herein, “cell-free extract” refers to an extract of thevegetative cells, spores and/or the whole culture broth of amicroorganism comprising cellular metabolites produced by the respectivemicroorganism obtainable by cell disruption methods known in the artsuch as solvent-based (e. g. organic solvents such as alcohols sometimesin combination with suitable salts), temperature-based, application ofshear forces, cell disruption with an ultrasonicator. The desiredextract may be concentrated by conventional concentration techniquessuch as drying, evaporation, centrifugation or alike. Certain washingsteps using organic solvents and/or water-based media may also beapplied to the crude extract preferably prior to use.

As used herein, the term “metabolite” refers to any component, compound,substance or byproduct (including but not limited to small moleculesecondary metabolites, polyketides, fatty acid synthase products,non-ribosomal peptides, ribosomal peptides, proteins and enzymes)produced by a microorganism (such as fungi and bacteria, in particularthe strains of the invention) that has any beneficial effect asdescribed herein such as pesticidal activity or improvement of plantgrowth, water use efficiency of the plant, plant health, plantappearance, or the population of beneficial microorganisms in the soilaround the plant activity herein.

As used herein, “isolate” refers to a pure microbial culture separatedfrom its natural origin, such an isolate obtained by culturing a singlemicrobial colony. An isolate is a pure culture derived from aheterogeneous, wild population of microorganisms.

As used herein, “strain” refers to a bacterial isolate or a group ofisolates exhibiting phenotypic and/or genotypic traits belonging to thesame lineage, distinct from those of other isolates or strains of thesame species.

As used herein, “a culture medium thereof” refers to a culture medium ofthe bacterial strain as defined right before the term “a culture mediumthereof”, as in the following case: “strains Lu16774, Lu17007 andLu17015, and a culture medium thereof” means the strains Lu16774,Lu17007 and Lu17015 and the culture medium of each of the strainsLu16774, Lu17007 and Lu17015. Likewise, the same logic applies tosimilar terms such as “a cell-free extract thereof”, “whole culturebroth thereof” and “metabolite thereof” as well as as combinations ofsuch terms such as “a cell-free extract or at least one metabolitethereof”.

A further embodiment relates to mixtures comprising as component 1) atleast one Paenibacillus strain having the following characteristics:

-   -   rod-shaped cells of Gram-positive structure,    -   weak reaction with Gram's stain, often even stain negatively,    -   differentiation into ellipsoidal endospores which distinctly        swell the sporangium (mother cell),    -   facultative anaerobic growth with strong growth in absence of        air irrespective of whether nitrate is present or not,    -   fermentation of a variety of sugars,    -   acid and gas formation from various sugars including glucose,    -   no acid production from adonitol and sorbitol,    -   Urease-negative (with exception of P. validus),    -   arginine dihydrolase negative,    -   no utilization of citrate,    -   no growth in presence of 10% sodium chloride,    -   secretion of numerous extracellular hydrolytic enzymes degrading        DNA, protein, starch; and/or    -   G+C content of DNA from 40% to 54%;        or a culture medium, a cell-free extract or at least one        metabolite thereof.

A further embodiment relates to mixtures comprising as component 1) atleast one Paenibacillus strain selected from the species Paenibacilluspolymyxa, Paenibacillus epiphyticus, Paenibacillus peoriae,Paenibacillus terrae, Paenibacillus jamilae, Paenibacillus kribbensis,Paenibacillus amylolyticus, Paenibacillus barcinonensis, Paenibacillustundra, Paenibacillus illinoisensis, Paenibacillus macquariensis,Paenibacillus taichungensis, Paenibacillus glycanilyticus andPaenibacillus odorifer, or a culture medium, a cell-free extract or atleast one metabolite of thereof.

A further embodiment relates to mixtures comprising as component 1) atleast one Paenibacillus strain selected from the species Paenibacilluspolymyxa, Paenibacillus epiphyticus, Paenibacillus peoriae,Paenibacillus terrae, Paenibacillus jamilae and Paenibacilluskribbensis, or a culture medium, a cell-free extract or at least onemetabolite thereof.

A further embodiment relates to mixtures comprising as component 1) atleast one bacterial strain selected from the species Paenibacilluspolymyxa, Paenibacillus epiphyticus, Paenibacillus peoriae andPaenibacillus jamilae, or a culture medium, a cell-free extract or atleast one metabolite thereof.

A further embodiment relates to mixtures comprising as component 1) atleast one bacterial strain selected from the species Paenibacilluspolymyxa, Paenibacillus epiphyticus and Paenibacillus peoriae, or aculture medium, a cell-free extract or at least one metabolite thereof.

A further embodiment relates to mixtures comprising as component 1) atleast one bacterial strain selected from the species Paenibacilluspolymyxa and Paenibacillus epiphyticus, or a culture medium, a cell-freeextract or at least one metabolite thereof.

A further embodiment relates to mixtures comprising as component 1) atleast one Paenibacillus strain selected from

1) Lu16774 deposited with DSMZ under Accession No. DSM 26969,

2) Lu17007 deposited with DSMZ under Accession No. DSM 26970, and

3) Lu17015 deposited with DSMZ under Accession No. DSM 26971;

or a culture medium, a cell-free extract or at least one metabolitethereof.

A further embodiment relates to mixtures comprising as component 1) atleast one Paenibacillus strain selected from

1) Paenibacillus polymyxa ssp. plantarum strain Lu16774 deposited withDSMZ under Accession No. DSM 26969,

2) Paenibacillus polymyxa ssp. plantarum strain Lu17007 deposited withDSMZ under Accession No. DSM 26970, and

3) Paenibacillus epiphyticus strain Lu17015 deposited with DSMZ underAccession No. DSM 26971;

or a culture medium, a cell-free extract or at least one metabolitethereof.

In addition to mixtures comprising as component 1) at least one of thestrains Lu16774, Lu17007 and Lu17015, the invention relates to mixturescomprising as component 1) any Paenibacillus strain, whether physicallyderived from the original deposit of any of the strains Lu16774, Lu17007and Lu17015 or independently isolated, so long as they retain at leastone of the identifying characteristics of the deposited Paenibacillusstrains Lu16774, Lu17007 and Lu17015. Such Paenibacillus strains of theinvention include any progeny of any of the strains Lu16774, Lu17007 andLu17015, including mutants of said strains.

The term “mutant” refers a microorganism obtained by direct mutantselection but also includes microorganisms that have been furthermutagenized or otherwise manipulated (e. g., via the introduction of aplasmid). Accordingly, embodiments include mutants, variants, and orderivatives of the respective microorganism, both naturally occurringand artificially induced mutants. For example, mutants may be induced bysubjecting the microorganism to known mutagens, such as X-ray, UVradiation or N-methyl-nitrosoguanidine, using conventional methods.Subsequent to said treatments a screening for mutant strains showing thedesired characteristics may be performed.

Mutant strains may be obtained by any methods known in the art such asdirect mutant selection, chemical mutagenesis or genetic manipulation(e. g., via the introduction of a plasmid). For example, such mutantsare obtainable by applying a known mutagen, such as X-ray, UV radiationor N-methyl-nitrosoguanidine. Subsequent to said treatments a screeningfor mutant strains showing the desired characteristics may be performed.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a DNAsequence exhibiting at least at least 99.6%, preferably at least 99.8%,even more preferably at least 99.9%, and in particular 100.0% nucleotidesequence identity to any one of the 16S rDNA sequences of the strainsLu16774, Lu17007 and Lu17015, i.e. to any one of those nucleotidesequences set forth in the Sequence listing being SEQ ID NO:1, SEQ IDNO:2 and SEQ ID NO:3.

According to a further embodiment, a mixture of the invention is inparticular one which comprises as component 1) at least one bacterialstrain which comprises a DNA sequence exhibiting 100% nucleotidesequence identity to any one of the 16S rDNA sequences of the strainsLu16774, Lu17007 and Lu17015, i.e. to any one of those nucleotidesequences set forth in the Sequence listing being SEQ ID NO:1, SEQ IDNO:2 or SEQ ID NO:3.

According to a further embodiment, a mixture of the invention is inparticular one which comprises as component 1) at least one bacterialstrain whose complete 16S rDNA sequence has after optimal alignmentwithin the aligned sequence window at least 99.6% identity to at leastone of the sequences SEQ ID NO:1 and SEQ ID NO:2 or at least 99.8%identity to SEQ ID NO:3; preferably at least 99.8% identity to at leastone of the sequences SEQ ID NO:1, SEQ ID:2 and SEQ ID NO:3; morepreferably at least 99.9% identity to at least one of the sequences SEQID NO:1, SEQ ID NO:2 and SEQ ID NO:3; even more preferably greater than99.9% identity to at least one of the sequences SEQ ID NO:1, SEQ ID:2and SEQ ID NO:3; in particular 100% identity to at least one of thesequences SEQ ID NO:1, SEQ ID:2 and SEQ ID NO:3.

According to a further embodiment, a mixture of the invention is inparticular one which comprises as component 1) at least one bacterialstrain selected from the group consisting of:

-   -   a) Lu16774 deposited with DSMZ under Accession No. DSM 26969;    -   b) Lu17007 deposited with DSMZ under Accession No. DSM 26970;    -   c) Lu17015 deposited with DSMZ under Accession No. DSM 26971;        and    -   d) a strain which comprises a DNA sequence exhibiting        -   d1) at least 99.6% nucleotide sequence identity to the DNA            sequences SEQ ID NO:4 or SEQ ID NO:9; or        -   d2) at least 99.8% nucleotide sequence identity to the DNA            sequence SEQ ID NO:14; or        -   d3) at least 99.9% nucleotide sequence identity to the DNA            sequences SEQ ID NO:5 or SEQ ID NO:10; or        -   d4) at least 99.2% nucleotide sequence identity to the DNA            sequence SEQ ID NO:15; or        -   d5) at least 99.2% nucleotide sequence identity to the DNA            sequences SEQ ID NO:6 or SEQ ID NO:11; or        -   d6) at least 99.8% nucleotide sequence identity to the DNA            sequence SEQ ID NO:16; or        -   d7) at least 99.8% nucleotide sequence identity to the DNA            sequences SEQ ID NO:7 or SEQ ID NO:12; or        -   d8) at least 99.3% nucleotide sequence identity to the DNA            sequence SEQ ID NO:17; or        -   d9) 100.0% nucleotide sequence identity to the DNA sequences            SEQ ID NO:8 or SEQ ID NO:13; or        -   d10) at least 99.9% nucleotide sequence identity to the DNA            sequence SEQ ID NO:18.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a dnaN DNAsequence exhibiting at least 99.6% nucleotide sequence identity to theDNA sequences SEQ ID NO:4 or SEQ ID NO:9 or which comprises a DNAsequence exhibiting at least 99.8% nucleotide sequence identity to theDNA sequence SEQ ID NO:14.

According to a further embodiment, a mixture of the invention is onewhich comprises as component 1) at least one bacterial strain whosecomplete dnaN DNA sequence has after optimal alignment within thealigned sequence window at least 99.6% identity to at least one of theDNA sequences SEQ ID NO:4 and SEQ ID NO:9 or at least 99.8% identity toSEQID NO:14; preferably at least 99.9% identity to SEQ ID NO:14; inparticular 100% identity to SEQ ID NO:14.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a DNAsequence exhibiting at least 99.8%, in particular 100.0% nucleotidesequence identity to any one of the dnaN DNA sequences of the strainsLu16774, Lu17007 and Lu17015, i.e. to any one of those DNA sequences SEQID NO:4, SEQ ID NO:9 and SEQ ID NO:14.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a gyrB DNAsequence exhibiting at least 99.9% nucleotide sequence identity to theDNA sequences SEQ ID NO:5 or SEQ ID NO:10 or which comprises a DNAsequence exhibiting at least 99.2% nucleotide sequence identity to theDNA sequence SEQ ID NO:15.

According to a further embodiment, a mixture of the invention is onewhich comprises as component 1) at least one bacterial strain whosecomplete gyrB DNA sequence has after optimal alignment within thealigned sequence window at least 99.9% identity to at least one of theDNA sequences SEQ ID NO:5 and SEQ ID NO:10 or at least 99.9% identity toSEQID NO:15; preferably at least 99.9% identity to SEQ ID NO:15; inparticular 100% identity to SEQ ID NO:15.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a DNAsequence exhibiting 100.0% nucleotide sequence identity to any one ofthe gyrB DNA sequences of the strains Lu16774, Lu17007 and Lu17015, i.e.to any one of those DNA sequences SEQ ID NO:5, SEQ ID NO:10 and SEQ IDNO:15.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a recF DNAsequence exhibiting at least 99.2% nucleotide sequence identity to theDNA sequences SEQ ID NO:6 or SEQ ID NO:11 or which comprises a DNAsequence exhibiting at least 99.8% nucleotide sequence identity to theDNA sequence SEQ ID NO:16.

According to a further embodiment, a mixture of the invention is onewhich comprises as component 1) at least one bacterial strain whosecomplete recF DNA sequence has after optimal alignment within thealigned sequence window at least 99.2% identity to at least one of theDNA sequences SEQ ID NO:6 and SEQ ID NO:11 or at least 99.8% identity toSEQID NO:16; preferably at least 99.9% identity to SEQ ID NO:16; inparticular 100% identity to SEQ ID NO:16.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a DNAsequence exhibiting at least 99.8%, in particular 100.0% nucleotidesequence identity to any one of the recFDNA sequences of the strainsLu16774, Lu17007 and Lu17015, i.e. to any one of those DNA sequences SEQID NO:6, SEQ ID NO:11 and SEQ ID NO:16.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a recti DNAsequence exhibiting at least 99.8% nucleotide sequence identity to theDNA sequences SEQ ID NO:7 or SEQ ID NO:12 or which comprises a DNAsequence exhibiting at least 99.3% nucleotide sequence identity to theDNA sequence SEQ ID NO:17.

According to a further embodiment, a mixture of the invention is onewhich comprises as component 1) at least one bacterial strain whosecomplete recN DNA sequence has after optimal alignment within thealigned sequence window at least 99.8% identity to at least one of theDNA sequences SEQ ID NO:7 and SEQ ID NO:12 or at least 99.3% identity toSEQID NO:17; preferably at least 99.6% identity to SEQ ID NO:17; inparticular 100% identity to SEQ ID NO:17.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a DNAsequence exhibiting at least 99.8%, in particular 100.0% nucleotidesequence identity to any one of the recti DNA sequences of the strainsLu16774, Lu17007 and Lu17015, i.e. to any one of those DNA sequences SEQID NO:7, SEQ ID NO:12 and SEQ ID NO:17.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a rpoA DNAsequence exhibiting 100.0% nucleotide sequence identity to the DNAsequences SEQ ID NO:8 or SEQ ID NO:13 or which comprises a DNA sequenceexhibiting at least 99.9% nucleotide sequence identity to the DNAsequence SEQ ID NO:18.

According to a further embodiment, a mixture of the invention is onewhich comprises as component 1) at least one bacterial strain whosecomplete rpoA DNA sequence has after optimal alignment within thealigned sequence window 100.0% identity to at least one of the DNAsequences SEQ ID NO:8 and SEQ ID NO:13 or at least 99.9% identity toSEQID NO:18; preferably at least 99.9% identity to SEQ ID NO:17; inparticular 100% identity to SEQ ID NO:18.

A mixture of the invention is in particular one which comprises ascomponent 1) at least one bacterial strain which comprises a DNAsequence exhibiting 100.0% nucleotide sequence identity to any one ofthe rpoA DNA sequences of the strains Lu16774, Lu17007 and Lu17015, i.e.to any one of those DNA sequences SEQ ID NO:8, SEQ ID NO:13 and SEQ IDNO:18.

A further embodiment relates to a mixture which comprises ascomponent 1) at least one isolated microorganism, being a member of thegenus Paenibacillus, having at least one of the identifyingcharacteristics of one of the following strains:

1) Paenibacillus strain Lu16774 deposited with DSMZ under Accession No.DSM 26969,

2) Paenibacillus strain Lu17007 deposited with DSMZ under Accession No.DSM 26970, or

3) Paenibacillus strain Lu17015 deposited with DSMZ under Accession No.DSM 26971.

A further embodiment relates to a mixture which comprises ascomponent 1) at least one Paenibacillus strain, which is selected fromthe group consisting of:

1) strain Lu16774 deposited with DSMZ under Accession No. DSM 26969,

2) strain Lu17007 deposited with DSMZ under Accession No. DSM 26970,

3) strain Lu17015 deposited with DSMZ under Accession No. DSM 26971, and

4) strains having at least one of the identifying characteristics of oneof said strains Lu16774, Lu17007 and Lu17015.

A further embodiment relates to a mixture which comprises ascomponent 1) at least one isolated microorganism selected from strains:

1) Paenibacillus strain Lu16774 deposited with DSMZ under Accession No.DSM 26969,

2) Paenibacillus strain Lu17007 deposited with DSMZ under Accession No.DSM 26970, and

3) Paenibacillus strain Lu17015 deposited with DSMZ under Accession No.DSM 26971; showing antagonistic activity against at least one plantpathogen, and being capable of producing at least one fusaricidin; or amutant strain thereof retaining said capability, i.e. retaining saidantagonistic activity against at least one plant pathogen, and retainingsaid capability of producing at least one fusaricidin.

A further embodiment relates to a mixture which comprises ascomponent 1) at least one microorganism selected from:

1) Paenibacillus strain Lu16774 deposited with DSMZ under Accession No.DSM 26969,

2) Paenibacillus strain Lu17007 deposited with DSMZ under Accession No.DSM 26970,

3) Paenibacillus strain Lu17015 deposited with DSMZ under Accession No.DSM 26971;

or a mutant strain thereof having all the identifying characteristics ofone of said strains.

A further embodiment relates to mixtures comprising as component 1) theculture medium of at least one Paenibacillus strain as defined in anyone of the preferred embodiments above.

A further embodiment relates to mixtures comprising as component 1) thecell-free extract of at least one Paenibacillus strain as defined in anyone of the preferred embodiments above.

A further embodiment relates to mixtures comprising as component 1) atleast one metabolite of at least Paenibacillus strain as defined in anyone of the preferred embodiments above; preferably the at least onemetabolite being a lipopeptide and even more preferably selected thegroups of polymyxins, octapeptins, polypeptins, pelgipeptins andfusaricidins.

A further embodiment relates to mixtures comprising as component 1) atleast one fusaricidin.

A further embodiment relates to mixtures comprising as component 1) atleast one fusaricidin of formula I

whereinR is selected from 15-guanidino-3-hydroxypentadecanoic acid (GHPD) and12-guanidinododecanoic acid (12-GDA);X¹ is threonine;X² is selected from isoleucine and valine;X³ is selected from tyrosine, valine, isoleucine and phenylalanine;X⁴ is threonine;X⁵ is selected from glutamine and asparagine;X⁶ is alanine; andwherein an arrow defines a single (amide) bond either between thecarbonyl moiety of R and the amino group of the amino acid X¹ or betweenthe carbonyl group of one amino acid and the amino group of aneighboring amino acid, wherein the tip of the arrow indicates theattachment to the amino group of said amino acid X¹ or of saidneighboring amino acid; andwherein the single line without an arrow head defines a single (ester)bond between the carbonyl group of X⁶ and the hydroxyl group of X¹.

A further embodiment relates to mixtures comprising as component 1) atleast one fusaricidin of forumula I.1

whereinR is selected from 15-guanidino-3-hydroxypentadecanoic acid (GHPD) and12-guanidinododecanoic acid (12-GDA);X¹ is threonine;X² is isoleucine;X³ is tyrosine;X⁴ is threonine;X⁵ is selected from glutamine and asparagine;X⁶ is alanine; andwherein an arrow defines a single (amide) bond either between thecarbonyl moiety of R and the amino group of the amino acid X¹ or betweenthe carbonyl group of one amino acid and the amino group of aneighboring amino acid, wherein the tip of the arrow indicates theattachment to the amino group of said amino acid X¹ or of saidneighboring amino acid; andwherein the single line without an arrow head defines a single (ester)bond between the carbonyl group of X⁶ and the hydroxyl group of X¹.

According to a further embodiment, X¹ in formula I is preferablyL-threonine. According to a further embodiment, X² in formula I ispreferably D-isoleucine or D-allo-isoleucine. According to a furtherembodiment, X³ in formula I is preferably L-tyrosine. According to afurther embodiment, X⁴ in formula I is preferably D-allo-threonine.According to a further embodiment, X⁵ in formula I is preferablyD-glutamine or D-asparagine. According to a further embodiment, R informula I is preferably GHPD.

The sketch of formula I.1 for fusaricidin of formula I.1 may also bedepicted as follows:

whereinX is selected from —NH—(C═O)—CH₂—CH(OH)—(CH₂)₁₂—NH—C(═NH)NH₂ and—NH—(C═O)—(CH₂)₁₁—NH—C(═NH)NH₂;R¹ is 1-hydroxyethyl;R² is 1-methylpropyl (sec-butyl);R³ is 4-hydroxybenzyl;R⁴ is 1-hydroxyethyl;R⁵ is selected from carbamoylethyl and carbamoylmethyl;R⁶ is methyl.

Likewise, the preferred embodiments based on the abovementionedalternative sketch of formula I.1 are as follows:

R¹ in this formula I is preferably (1S,2R)-1-hydroxyethyl.

R² in this formula I is preferably (1R,2R)-1-methylpropyl or(1R,2S)-1-methylpropyl.

R³ in this formula I is preferably (S)-4-hydroxy-benzyl.

R⁴ in this formula I is preferably (1S,2R)-1-hydroxyethyl.

R⁵ in this formula I is preferably (R)-carbamoylethyl and(R)-carbamoylmethyl.

X in this formula I is preferably—NH—(C═O)—CH₂—CH(OH)—(CH₂)₁₂—NH—C(═NH)NH₂.

According to a further embodiment, the invention further relates tomixtures comprising as component 1) at east one fusaricidin selectedfrom fusaricidin 1A and 1B, which are of formula I, wherein R is GHPDand wherein X⁵ is asparagine in case of fusaricidin 1A and X⁵ isglutamine in case of fusaricidin 1B:

Further, the fusaricidins of formula I including those wherein R is GHTDcan be synthesized in analogy to methods known in the art (Biopolymers80(4), 541, 2005; J. Peptide Sci. 12S, 219, 2006; Tetrahedron Lett.47(48), 8587-90, 2006; Biopolymers 88(4), 568, 2007; ChemMedChem 7,871-882, 2012).

The present invention further relates to compositions comprising themixtures of the inventions which comprise as component 1) the strains,whole culture broth, cell-free extracts, culture media, or fusaricidinsof formula I and their salts of the invention, as well as to the use ofsaid compositions for controlling or suppressing plant pathogens orpreventing plant pathogen infection or for protection of materialsagainst infestation destruction by harmful microorganisms, and tocorresponding methods which comprise treating the pathogens, theirhabitat or the materials or plants to be protected against pathogenattack, or the soil or propagation material with an effective amount ofthe compositions, strains, whole culture broth, cell-free extracts,culture media, or fusaricidins of formula I and their salts of theinvention.

Further embodiments of the invention are disclosed in the followingdetailed description of the invention, the claims and the figures.

An identifying characteristic of the deposited Paenibacillus strainsLu16774, Lu17007 and Lu17015 is that they are capable of producing atleast one fusaricidin of formula I, preferably selected fromfusaricidins 1A and 1B, in particular producing fusaricidins 1A and 1B,which are metabolites of the respective strains.

Thus, according to one aspect of the invention, the Paenibacillusstrains of component 1) of the mixtures of the invention are capable ofproducing at least one fusaricidin of formula I, more preferablyproducing fusaricidins 1A or 1B, in particular producing fusaricidins 1Aand 1B; more preferably in a growth medium comprising at least onesource of carbon and one source of nitrogen as defined herein.

Thus, according to another aspect of the invention, Paenibacillusstrains of component 1) of the mixtures of the invention produce atleast one fusaricidin of formula I, more preferably produce fusaricidins1A or 1B, in particular produce fusaricidins 1A and 1B; in a growthmedium comprising at least one source of carbon and one source ofnitrogen as defined herein.

Another embodiment of the invention relates to mixtures comprising ascomponent 1) at least one isolated microorganism selected from

1) Paenibacillus strain Lu16774 deposited with DSMZ under Accession No.DSM 26969,

2) Paenibacillus strain Lu17007 deposited with DSMZ under Accession No.DSM 26970, and

3) Paenibacillus strain Lu17015 deposited with DSMZ under Accession No.DSM 26971;

showing antagonistic activity against at least one plant pathogen, andbeing capable of producing at least one fusaricidin of formula I,preferably selected from fusaricidins 1A and 1B, in particular producingfusaricidins 1A and 1B; or a mutant strain thereof retaining saidcapability, i.e. retaining said antagonistic activity against at leastone plant pathogen, and retaining said capability of producing at leastone fusaricidin of formula I, preferably selected from fusaricidins 1Aand 1B, in particular producing fusaricidins 1A and 1B.

A further identifying characteristic of the Paenibacillus strainsLu16774, Lu17007 and Lu17015 or a mutant strain thereof is that they arecapable of producing at least one fusaricidin selected from the groupconsisting of fusaricidin A, fusaricidin B, fusaricidin C, fusaricidinD, LI-F06a, LI-F06b and LI-F08b in addition to their capability ofproducing at least one fusaricidin of formula I, preferably selectedfrom fusaricidin 1A and 1B, in particular producing fusaricidin 1A and1B.

Thus, according to a further aspect of the invention, Paenibacillusstrains of component 1) of the mixtures of the invention are capable ofproducing at least one fusaricidin of formula I, preferably selectedfrom fusaricidins 1A and 1B, in particular producing fusaricidins 1A and1B, as disclosed herein, and are capable of producing at least onecompound selected from the group consisting of fusaricidin A,fusaricidin B, fusaricidin C, fusaricidin D, LI-F06a, LI-F06b andLI-F08b.

According to a further aspect of the invention, Paenibacillus strains ofcomponent 1) of the mixtures of the invention are capable of producingat least one fusaricidin of formula I, preferably selected fromfusaricidins 1A and 1 B, in particular producing fusaricidins 1A and 1B,as disclosed herein, and are capable of producing at least threecompounds selected from the group consisting of fusaricidin A,fusaricidin B, fusaricidin C, fusaricidin D, LI-F06a, LI-F06b andLI-F08b.

A further identifying characteristic of the Paenibacillus strains aretheir antifungal activity. In particular, these strains were found to beeffective against infestion with plant pathogens including Alternariaspp., Botrytis cinerea, Phytophthora infestans, and Sclerotiniasclerotiorum; wherein Alternaria spp. is preferably selected from A.solani and A. alternata, in particular A. solani.

Thus, according to a further aspect of the invention, Paenibacillusstrains of component 1) of the mixtures of the invention have antifungalactivity, particularly against a plant pathogen selected from the groupconsisting of Alternaria spp., Botrytis cinerea, Phytophthora infestans,and Sclerotinia sclerotiorum, wherein Alternaria spp. is preferablyselected from A. solani and A. altemata, in particular A. solani. Moreparticularly, Paenibacillus strains of the invention have antifungalactivity against at least two or against all four of said pathogens.

According to a further aspect of the invention, Paenibacillus strains ofthe invention have antifungal activity against the plant pathogensAlternaria solani, Botrytis cinerea, Phytophthora infestans, andSclerotinia sclerotiorum.

Antagonistic activity of the Paenibacillus strains against plantpathogens can be shown in an in-vitro confrontation assays using thedesired phytopathogenic fungi such as Alternaria spp., Botrytis cinerea,Phytophthora infestans, and Sclerotinia sclerotiorum wherein Alternariaspp. is preferably selected from A. solani and A. alternata, inparticular A. solani.

As growth medium for these phytopathogenic fungi, ISP2 medium is usedcomprising per litre: 10 g malt extract (Sigma Aldrich, 70167); 4 gBacto yeast extract (Becton Dickinson, 212750); 4 g glucose monohydrate(Sigma Aldrich, 16301); 20 g Agar (Becton Dickinson, 214510), pH about7, Aq. bidest. As growth medium for PHYTIN, V8 medium is used comprisingper litre: 200 ml of vegetable juice, 3 g calcium carbonate (MerckMillipore, 1020660250); 30 g Agar (Becton Dickinson, 214510), pH 6.8,Aq. bidest.

The Paenibacillus strains are point-inoculated on one side of an agarplate. An agar block (approx. 0.3 cm²) containing one actively growingplant pathogen was put in the center of the plate. After incubating for7-14 days at about 25° C., the growth of the plant pathogen is examined,especially for inhibition zones. The following antagonistic effects canbe evaluated: Antibiosis is scored by evaluation of the diameter of thefungi-free zone (zone of inhibition). Competition is scored by comparingthe diameter of the growth of the fungal pathogen on plates withbacterial strains in comparison to control plates. Mycoparasitism can bedocumented in case the bacteria overgrows the fungal pathogen and alsomycoparasite the pathogens. This can be visualized by microscopy.

More specifically, the present invention relates mixtures comprising ascomponent 1) at least one bacterial strain selected from Paenibacillusstrains Lu16774, Lu17007 and Lu17015 and any Paenibacillus strain havingone, two, three or more of the identifying characteristics of thedeposited strain, wherein the identifying characteristics are selectedfrom the group consisting of:

(a) antifungal activity against a plant pathogen selected from the groupconsisting of Alternaria spp., Botrytis cinerea, Phytophthora infestans,and Sclerotinia sclerotiorum, wherein Alternaria spp. is preferablyselected from A. solani and A. alternata, in particular A. solani, asdisclosed herein;

(b) the capability of producing at least one fusaricidin of formula I,in particular fusaricidins 1A and/or 1B, as disclosed herein;

(c) the capability of producing at least one fusaricidin selected fromthe group consisting of fusaricidins A, B, C, D, LI-F06a, LI-F06b andLI-F08b, as disclosed herein; and

(d) the capability of producing and secreting at least one lytic enzymeselected from the group consisting of chitinase, cellulose and amylase,as disclosed herein.

-   -   More preferably, said Paenibacillus strain has the capabilities        referred to as (b), (c) and (d) in a growth medium comprising at        least one source of carbon and one source of nitrogen as defined        herein.

In particular, Paenibacillus strains of component 1) of the mixtures ofthe invention have two or more of the identifying characteristics of thedeposited strain, with strains having at least the characteristics (a)and (b) being particularly preferred. For instance, according to apreferred embodiment, the Paenibacillus strains of component 1) of themixtures of the invention (a) have an antifungal activity against aplant pathogen selected from the group consisting of Alternaria spp.,Botrytis cinerea, Phytophthora infestans, and Sclerotinia sclerotiorum,wherein Alternaria spp. is preferably selected from A. solani and A.alternata, in particular A. solani and (b) are capable of producing atleast one fusaricidin of formula I, and particularly fusaricidin 1B.According to a further preferred embodiment, the Paenibacillus strainsof component 1) of the mixtures of the invention (a) have an antifungalactivity against three or against all of the plant pathogens selectedfrom the group consisting of Alternaria spp., Botrytis cinerea,Phytophthora infestans, and Sclerotinia sclerotiorum, wherein Alternariaspp. is preferably selected from A. solani and A. alternata, inparticular A. solani and (b) are capable of producing at least onefusaricidin of formula I, more preferably producing fusaricidin 1A or1B, in particular of producing fusaricidin 1A and 1B.

According to an embodiment of the invention, the Paenibacillus strainsof component 1) of the mixtures of the invention are provided inisolated or substantially purified form.

The terms “isolated” or “substantially purified” are meant to denotethat the strains of the invention have been removed from a naturalenvironment and have been isolated or separated, and are at least 60%free, preferably at least 75% free, and more preferably at least 90%free, even more preferably at least 95% free, and most preferably atleast 99% free from other components with which they were naturallyassociated. An isolate obtained by culturing a single microbial colonyis an example of an isolated strain of the invention.

The mixtures of the invention comprise as component 1) the at least onePaenibacillus strain in any physiological state such as active ordormant. Dormant Paenibacillus strains may be provided for examplefrozen, dried, or lyophilized or partly desiccated (procedures toproduce partly desiccated organisms are given in WO 2008/002371) or inform of spores.

According to an embodiment of the invention, the mixtures comprise ascomponent 1) the at least one Paenibacillus strain in form of spores.

According to an embodiment of the invention, the mixtures comprise ascomponent 1) the at least one Paenibacillus strain in form of a wholeculture broth.

The culture is preferably an isolated or substantially purified culture.

An “isolated culture” or “substantially purified culture” refers to aculture of the strains of the invention that does not includesignificant amounts of other materials which normally are found innatural habitat in which the strain grows and/or from which the strainnormally may be obtained. Consequently, such “isolated culture” or“substantially purified culture” is at least 60% free, preferably atleast 75% free, and more preferably at least 90% free, even morepreferably at least 95% free, and most preferably at least 99% free fromother materials which normally are found in natural habitat in which thestrain grows and/or from which the strain normally may be obtained. Suchan “isolated culture” or “substantially purified culture” does normallynot include any other microorganism in quantities sufficient tointerfere with the replication of the strain of the invention.

The Paenibacillus strains as used in the mixtures of the invention canbe cultivated continuously or discontinuously in the batch process or inthe fed batch or repeated fed batch process. A review of known methodsof cultivation will be found in the textbook by Chmiel(Bioprozesstechnik 1. Einführung in die Bioverfahrenstechnik (GustavFischer Verlag, Stuttgart, 1991)) or in the textbook by Storhas(Bioreaktoren and periphere Einrichtungen (Vieweg Verlag,Braunschweig/Wiesbaden, 1994)).

The medium that is to be used for cultivation of the microorganism mustsatisfy the requirements of the particular strains in an appropriatemanner. Descriptions of culture media for various microorganisms aregiven in the handbook “Manual of Methods for General Bacteriology” ofthe American Society for Bacteriology (Washington D.C., USA, 1981).

These media that can be used according to the invention generallycomprise one or more sources of carbon, sources of nitrogen, inorganicsalts, vitamins and/or trace elements. Preferred sources of carbon aresugars, such as mono-, di- or polysaccharides. Very good sources ofcarbon are for example glucose, fructose, mannose, galactose, ribose,sorbose, ribulose, lactose, maltose, sucrose, raffinose, starch orcellulose. Sugars can also be added to the media via complex compounds,such as molasses, or other by-products from sugar refining. It may alsobe advantageous to add mixtures of various sources of carbon. Otherpossible sources of carbon are oils and fats such as soybean oil,sunflower oil, peanut oil and coconut oil, fatty acids such as palmiticacid, stearic acid or linoleic acid, alcohols such as glycerol, methanolor ethanol and organic acids such as acetic acid or lactic acid. Sourcesof nitrogen are usually organic or inorganic nitrogen compounds ormaterials containing these compounds. Examples of sources of nitrogeninclude ammonia gas or ammonium salts, such as ammonium sulfate,ammonium chloride, ammonium phosphate, ammonium carbonate or ammoniumnitrate, nitrates, urea, amino acids or complex sources of nitrogen,such as corn-steep liquor, soybean flour, soybean protein, yeastextract, meat extract and others. The sources of nitrogen can be usedseparately or as a mixture. Inorganic salt compounds that may be presentin the media comprise the chloride, phosphate or sulfate salts ofcalcium, magnesium, sodium, cobalt, molybdenum, potassium, manganese,zinc, copper and iron. Inorganic sulfur-containing compounds, forexample sulfates, sulfites, dithionites, tetrathionates, thiosulfates,sulfides, but also organic sulfur compounds, such as mercaptans andthiols, can be used as sources of sulfur. Phosphoric acid, potassiumdihydrogenphosphate or dipotassium hydrogenphosphate or thecorresponding sodium-containing salts can be used as sources ofphosphorus. Chelating agents can be added to the medium, in order tokeep the metal ions in solution. Especially suitable chelating agentscomprise dihydroxyphenols, such as catechol or protocatechuate, ororganic acids, such as citric acid. The fermentation media usedaccording to the invention may also contain other growth factors, suchas vitamins or growth promoters, which include for example biotin,riboflavin, thiamine, folic acid, nicotinic acid, pantothenate andpyridoxine. Growth factors and salts often come from complex componentsof the media, such as yeast extract, molasses, corn-steep liquor and thelike. In addition, suitable precursors can be added to the medium. Theprecise composition of the compounds in the medium is strongly dependenton the particular experiment and must be decided individually for eachspecific case. Information on media optimization can be found in thetextbook “Applied Microbiol. Physiology, A Practical Approach” (Publ. P.M. Rhodes, P. F. Stanbury, IRL Press (1997) p. 53-73, ISBN 0 19 9635773). Growing media can also be obtained from commercial suppliers, suchas Standard 1 (Merck) or BHI (Brain heart infusion, DIFCO) etc.

Preferred growth media that can be used according to the inventioncomprise one or more sources of carbon selected from L-arabinose,N-acetyl-D-glucosamine, D-galactose, L-aspartaic acid, D-trehalose,D-mannose, glycerol, D-gluconic acid, D-xylose, D-mannitol, D-ribose,D-fructose, α-D-glucose, maltose, D-melibiose, thymidine,α-methyl-D-Galactoside, α-D-lactose, lactulose, sucrose, uridine,α-hydroxy glutaric acid-γ-lactone, β-methyl-D-glucoside, adonitol,maltotriose, 2-deoxyadenosine, adenosine, citric acid, mucic acid,D-cellobiose, inosine, L-serine, L-alanyl-glycine, D-galacturonic acid,α-cyclodextrin, β-cyclodextrin, dextrin, inulin, pectin, amygdalin,gentiobiose, lactitol, D-melezitose, α-methyl-D-glucoside,β-methyl-D-galactoside, β-methyl-D-xyloside, palatinose, D-raffinose,stachyose, turanose, γ-amino butyric acid, D-gluosamine, D-lactic acid,L-lysine, 3-hydroxy 2-butanone; and one or more sources of nitrogenselected from ammonia, nitrite, nitrate, L-alaninie, L-asparagine,L-aspartic acid, L-glutamic acid, L-glutamie, glycine, aminoacid dimes:Ala-Asp, AlaGln, Ala-Glu, Ala-His, Gly-Gln, Gly-Glu, Gly-Met, andMet-Ala; in particular nitrate. These media can be supplemented withinorganic salts and vitamins and/or trace elements. The strains arecapable to produce fusaricidins 1A and 1B in these growth media.

All components of the medium are sterilized, either by heating (20 minat 2.0 bar and 121° C.) or by sterile filtration. The components can besterilized either together, or if necessary separately. All thecomponents of the medium can be present at the start of growing, oroptionally can be added continuously or by batch feed.

The temperature of the culture is normally between 15° C. and 36° C.,preferably 25° C. to 33° C. and can be kept constant or can be variedduring the experiment. The pH value of the medium should be in the rangefrom 5 to 8.5, preferably around 7.0. The pH value for growing can becontrolled during growing by adding basic compounds such as sodiumhydroxide, potassium hydroxide, ammonia or ammonia water or acidcompounds such as phosphoric acid or sulfuric acid. Antifoaming agents,e. g. fatty acid polyglycol esters, can be used for controlling foaming.To maintain the stability of plasmids, suitable substances withselective action, e. g. antibiotics, can be added to the medium. Oxygenor oxygen-containing gas mixtures, e. g. the ambient air, are fed intothe culture in order to maintain aerobic conditions. The temperature ofthe culture is normally from 20° C. to 45° C. Culture is continued untila maximum of the desired product has formed. This is normally achievedwithin 10 hours to 160 hours.

In particular, the strains of the invention may be cultivated in amedium a variety of standard microbiology media such as Luria-BertaniBroth (LB), trypticase-soy broth (TSB), yeast extract/maltextract/glucose broth (YMG, ISP2) at 15° C. to 36° C. for 18 to 360 h inliquid media or in agar-solidified media on a petri dish. Aeration maybe necessary. The bacterial cells (vegetative cells and spores) can bewashed and concentrated (e. g. by centrifugation at temperatures ofabout 15 to 30° C. for about 15 min at 7,000×g).

The invention also relates to mixtures comprising as component 1) aculture medium obtainable by culturing at least one one Paenibacillusstrain as defined in any one of the preferred embodiments above in amedium and separating the medium from the culture broth (thus, remainingwhen cells grown in the medium are removed from the whole culturebroth), e. g., the supernatant of a whole culture broth, i.e., theliquid broth remaining when cells grown in broth and other debris areremoved by centrifugation, filtration, sedimentation, or other meanswell known in the art. The supernatant can be obtained e. g. bycentrifugation at temperatures of about 2 to 30° C. (more preferably attemperatures of 4 to 20° C.) for about 10 to 60 min (more preferablyabout 15 to 30 min) at about 5,000 to 20,000×g (more preferably at about15,000×g).

Such culture medium contains pesticidal metabolites which are producedby the cultured strain.

The invention also relates to mixtures comprising as component 1) acell-free extract obtainable from at least Paenibacillus strain asdefined in any one of the preferred embodiments above. To produce acell-free extract, the strain may be cultivated as described above. Thecells can be disrupted also by high-frequency ultrasound, by highpressure, e. g. in a French pressure cell, by osmolysis, by the actionof detergents, lytic enzymes or organic solvents, by means ofhomogenizers or by a combination of several of the methods listed. Theextraction can be carried out preferably with an organic solvent orsolvent mixture, more preferably an alcohol (e. g. methanol, ethanol,n-propanol, 2-propanol or alike), even more preferably with 2-propanol(e. g. in a 1:1 ratio to the culture volume). Phase separation may beenhanced by addition of salts such as NaCl. The organic phase can becollected and the solvent or solvent mixture may be removed byconventional distillation and/or drying followed by resuspension inmethanol and filtration.

Such extract contains pesticidal metabolites which are produced by thecultured strain.

Pesticidal metabolites that are specific to the strains of the inventionmay be recovered from such medium or extract according to conventionalmethods in particular when the strains of the invention have beencultivated as described above.

The methodology of the present invention can further include a step ofrecovering individual pesticidal metabolites.

The term “recovering” includes extracting, harvesting, isolating orpurifying the compound from culture media or cell-free extracts.Recovering the compound can be performed according to any conventionalisolation or purification methodology known in the art including, butnot limited to, treatment with a conventional resin (e. g., anion orcation exchange resin, non-ionic adsorption resin, etc.), treatment witha conventional adsorbent (e. g., activated charcoal, silicic acid,silica gel, cellulose, alumina, etc.), alteration of pH, solventextraction (e. g., with a conventional solvent such as an alcohol, ethylacetate, hexane and the like), distillation, dialysis, filtration,concentration, crystallization, recrystallization, pH adjustment,lyophilization and the like. For example the metabolites can berecovered from culture media by first removing the microorganisms. Theremaining broth is then passed through or over a cation exchange resinto remove unwanted cations and then through or over an anion exchangeresin to remove unwanted inorganic anions and organic acids.

Consequently, the invention also relates to a mixture comprising ascomponent 1) a whole culture broth of a microorganism comprising atleast one fusaricidins of formula I preferably selected fromfusaricidins 1A and 1B, in particular said whole culture broth comprisesfusaricidins 1A and 1B.

According to a further embodiment, the invention also relates to amixture comprising as component 1) a whole culture broth of amicroorganism Paenibacillus strain comprising at least one fusaricidinsof formula I, preferably selected from fusaricidins 1A and 1B, inparticular said whole culture broth comprises fusaricidins 1A and 1B.

Said fusaricidin-type metabolites are secreted into the culture mediumof the respective microorganism capable of producing it.

Consequently, the invention also relates to a mixture comprising ascomponent 1) a culture medium and/or a cell-free extract of amicroorganism comprising at least one fusaricidin of formula I,preferably selected from fusaricidins 1A and 1B, in particular saidculture medium and/or a cell-free extract comprises fusaricidins 1A and1B.

According to a further embodiment, the invention also relates to amixture comprising as component 1) a culture medium and/or a cell-freeextract of a microorganism of the genus Paenibacillus comprising atleast one fusaricidin of formula I, preferably selected fromfusaricidins 1A and 1B, in particular said culture medium and/or acell-free extract comprises fusaricidins 1A and 1B.

According to a further embodiment, the invention also relates to amixture comprising as component 1) a culture medium and/or a cell-freeextract of at least one Paenibacillus strain of the invention as definedin any one of the preferred embodiments above comprising at least onefusaricidin of formula I as defined above, preferably selected fromfusaricidins 1A and 1B, in particular said culture medium and/or acell-free extract comprises fusaricidins 1A and 1B.

The invention further relates to agrochemical compositions comprising anauxiliary as defined below and the mixture of the invention comprisingas copmponent 1) at least one bacterial strain, whole culture broth,cell-free extract, culture medium and/or fusaricidin of formula I, asdefined in any one of the preferred embodiments above, respectively.

As used herein, “composition” in reference to a product (microbialstrain, agent or formulation) of the present invention refers to acombination of ingredients, wherein “formulating” is the process ofusing a formula, such as a recipe, for a combination of ingredients, tobe added to form the formulation. Such composition is also referredherein to as formulation.

The mixtures comprising as component 1) the bacterial strains, wholeculture broths, cell-free extracts, culture media, fusaricidins offormula I as defined in any one of the preferred embodiments above, andas component 2) at least one biopesticide II as defined in any one ofthe preferred embodiments above; and compositions of the invention,respectively, are suitable as antifungal agents or fungicides. They aredistinguished by an outstanding effectiveness against a broad spectrumof phytopathogenic fungi, including soil-borne fungi, which deriveespecially from the classes of the Plasmodiophoromycetes,Peronosporomycetes (syn. Oomycetes), Chytridiomycetes, Zygomycetes,Ascomycetes, Basidiomycetes and Deuteromycetes (syn. Fungi imperfecti).Some are systemically effective and they can be used in crop protectionas foliar fungicides, fungicides for seed dressing and soil fungicides.Moreover, they are suitable for controlling harmful fungi, which interalia occur in wood or roots of plants.

The mixtures and compositions of the invention, respectively, areparticularly important in the control of a multitude of phytopathogenicfungi on various cultivated plants, such as cereals, e. g. wheat, rye,barley, triticale, oats or rice; beet, e. g. sugar beet or fodder beet;fruits, such as pomes, stone fruits or soft fruits, e. g. apples, pears,plums, peaches, almonds, cherries, strawberries, raspberries,blackberries or gooseberries; leguminous plants, such as lentils, peas,alfalfa or soybeans; oil plants, such as rape, mustard, olives,sunflowers, coconut, cocoa beans, castor oil plants, oil palms, groundnuts or soybeans; cucurbits, such as squashes, cucumber or melons; fiberplants, such as cotton, flax, hemp or jute; citrus fruit, such asoranges, lemons, grapefruits or mandarins; vegetables, such as spinach,lettuce, asparagus, cabbages, carrots, onions, tomatoes, potatoes,cucurbits or paprika; lauraceous plants, such as avocados, cinnamon orcamphor; energy and raw material plants, such as corn, soybean, rape,sugar cane or oil palm; corn; tobacco; nuts; coffee; tea; bananas; vines(table grapes and grape juice grape vines); hop; turf; sweet leaf (alsocalled Stevia); natural rubber plants or ornamental and forestry plants,such as flowers, shrubs, broad-leaved trees or evergreens, e. g.conifers; and on the plant propagation material, such as seeds, and thecrop material of these plants.

Preferably, the mixtures and compositions of the invention,respectively, are used for controlling a multitude of fungi on fieldcrops, such as potatoes sugar beets, tobacco, wheat, rye, barley, oats,rice, corn, cotton, soybeans, rape, legumes, sunflowers, coffee or sugarcane; fruits; vines; ornamentals; or vegetables, such as cucumbers,tomatoes, beans or squashes.

The term “plant propagation material” is to be understood to denote allthe generative parts of the plant such as seeds and vegetative plantmaterial such as cuttings and tubers (e. g. potatoes), which can be usedfor the multiplication of the plant. This includes seeds, roots, fruits,tubers, bulbs, rhizomes, shoots, sprouts and other parts of plants,including seedlings and young plants, which are to be transplanted aftergermination or after emergence from soil. These young plants may also beprotected before transplantation by a total or partial treatment byimmersion or pouring.

Preferably, treatment of plant propagation materials with the strains,whole culture broths, cell-free extracts culture media, fusaricidins offormula I; and compositions of the invention, respectively, is used forcontrolling a multitude of fungi on cereals, such as wheat, rye, barleyand oats; rice, corn, cotton and soybeans.

The term “cultivated plants” is to be understood as including plantswhich have been modified by breeding, mutagenesis or genetic engineeringincluding but not limiting to agricultural biotech products on themarket or in development (cf. http://cera-gmc.org/, see GM crop databasetherein). Genetically modified plants are plants, which genetic materialhas been so modified by the use of recombinant DNA techniques that undernatural circumstances cannot readily be obtained by cross breeding,mutations or natural recombination. Typically, one or more genes havebeen integrated into the genetic material of a genetically modifiedplant in order to improve certain properties of the plant. Such geneticmodifications also include but are not limited to targetedpost-translational modification of protein(s), oligo- or polypeptides e.g. by glycosylation or polymer additions such as prenylated, acetylatedor farnesylated moieties or PEG moieties.

Plants that have been modified by breeding, mutagenesis or geneticengineering, e. g. have been rendered tolerant to applications ofspecific classes of herbicides, such as auxin herbicides such as dicambaor 2,4-D; bleacher herbicides such as hydroxylphenylpyruvate dioxygenase(HPPD) inhibitors or phytoene desaturase (PDS) inhibitors; acetolactatesynthase (ALS) inhibitors such as sulfonyl ureas or imidazolinones;enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibitors, such asglyphosate; glutamine synthetase (GS) inhibitors such as glufosinate;protoporphyrinogen-IX oxidase inhibitors; lipid biosynthesis inhibitorssuch as acetyl CoA carboxylase (ACCase) inhibitors; or oxynil (i. e.bromoxynil or ioxynil) herbicides as a result of conventional methods ofbreeding or genetic engineering. Furthermore, plants have been maderesistant to multiple classes of herbicides through multiple geneticmodifications, such as resistance to both glyphosate and glufosinate orto both glyphosate and a herbicide from another class such as ALSinhibitors, HPPD inhibitors, auxin herbicides, or ACCase inhibitors.These herbicide resistance technologies are e. g. described in PestManagem. Sci. 61, 2005, 246; 61, 2005, 258; 61, 2005, 277; 61, 2005,269; 61, 2005, 286; 64, 2008, 326; 64, 2008, 332; Weed Sci. 57, 2009,108; Austral. J. Agricult. Res. 58, 2007, 708; Science 316, 2007, 1185;and references quoted therein. Several cultivated plants have beenrendered tolerant to herbicides by conventional methods of breeding(mutagenesis), e. g. Clearfield® summer rape (Canola, BASF SE, Germany)being tolerant to imidazolinones, e. g. imazamox, or ExpressSun®sunflowers (DuPont, USA) being tolerant to sulfonyl ureas, e. g.tribenuron. Genetic engineering methods have been used to rendercultivated plants such as soybean, cotton, corn, beets and rape,tolerant to herbicides such as glyphosate and glufosinate, some of whichare commercially available under the trade names RoundupReady®(glyphosate-tolerant, Monsanto, U.S.A.), Cultivance® (imidazolinonetolerant, BASF SE, Germany) and LibertyLink® (glufosinate-tolerant,Bayer CropScience, Germany).

Furthermore, plants are also covered that are by the use of recombinantDNA techniques capable to synthesize one or more insecticidal proteins,especially those known from the bacterial genus Bacillus, particularlyfrom Bacillus thuringiensis, such as δ-endotoxins, e. g. CryIA(b),CryIA(c), CryIF, CryIF(a2), CryIIA(b), CryIIIA, CryIIIB(b1) or Cry9c;vegetative insecticidal proteins (VIP), e. g. VIP1, VIP2, VIP3 or VIP3A;insecticidal proteins of bacteria colonizing nematodes, e. g.Photorhabdus spp. or Xenorhabdus spp.; toxins produced by animals, suchas scorpion toxins, arachnid toxins, wasp toxins, or otherinsect-specific neurotoxins; toxins produced by fungi, suchStreptomycetes toxins, plant lectins, such as pea or barley lectins;agglutinins; proteinase inhibitors, such as trypsin inhibitors, serineprotease inhibitors, patatin, cystatin or papain inhibitors;ribosome-inactivating proteins (RIP), such as ricin, maize-RIP, abrin,luffin, saporin or bryodin; steroid metabolism enzymes, such as3-hydroxysteroid oxidase, ecdysteroid-IDP-glycosyl-transferase,cholesterol oxidases, ecdysone inhibitors or HMG-CoA-reductase; ionchannel blockers, such as blockers of sodium or calcium channels;juvenile hormone esterase; diuretic hormone receptors (helicokininreceptors); stilbene synthase, bibenzyl synthase, chitinases orglucanases. In the context of the present invention these insecticidalproteins or toxins are to be understood expressly also as pre-toxins,hybrid proteins, truncated or otherwise modified proteins. Hybridproteins are characterized by a new combination of protein domains,(see, e. g. WO 02/015701). Further examples of such toxins orgenetically modified plants capable of synthesizing such toxins aredisclosed, e. g., in EP-A 374 753, WO 93/007278, WO 95/34656, EP-A 427529, EP-A 451 878, WO 03/18810 and WO 03/52073. The methods forproducing such genetically modified plants are generally known to theperson skilled in the art and are described, e. g. in the publicationsmentioned above. These insecticidal proteins contained in thegenetically modified plants impart to the plants producing theseproteins tolerance to harmful pests from all taxonomic groups ofarthropods, especially to beetles (Coeloptera), two-winged insects(Diptera), and moths (Lepidoptera) and to nematodes (Nematoda).Genetically modified plants capable to synthesize one or moreinsecticidal proteins are, e. g., described in the publicationsmentioned above, and some of which are commercially available such asYieldGard® (corn cultivars producing the CrylAb toxin), YieldGard® Plus(corn cultivars producing Cry1Ab and Cry3Bb1 toxins), Starlink® (corncultivars producing the Cry9c toxin), Herculex® RW (corn cultivarsproducing Cry34Ab1, Cry35Ab1 and the enzymePhosphinothricin-N-Acetyltransferase [PAT]); NuCOTN® 33B (cottoncultivars producing the CrylAc toxin), Bollgard® I (cotton cultivarsproducing the CrylAc toxin), Bollgard® II (cotton cultivars producingCrylAc and Cry2Ab2 toxins); VIPCOT° (cotton cultivars producing aVIP-toxin); NewLeaf® (potato cultivars producing the Cry3A toxin);Bt-Xtra®, NatureGard®, KnockOut®, BiteGard®, Protecta®, Bt11 (e. g.Agrisure® CB) and Bt176 from Syngenta Seeds SAS, France, (corn cultivarsproducing the CrylAb toxin and PAT enzyme), MIR604 from Syngenta SeedsSAS, France (corn cultivars producing a modified version of the Cry3Atoxin, c.f. WO 03/018810), MON 863 from Monsanto Europe S.A., Belgium(corn cultivars producing the Cry3Bb1 toxin), IPC 531 from MonsantoEurope S.A., Belgium (cotton cultivars producing a modified version ofthe CrylAc toxin) and 1507 from Pioneer Overseas Corporation, Belgium(corn cultivars producing the Cry1F toxin and PAT enzyme).

Furthermore, plants are also covered that are by the use of recombinantDNA techniques capable to synthesize one or more proteins to increasethe resistance or tolerance of those plants to bacterial, viral orfungal pathogens. Examples of such proteins are the so-called“pathogenesis-related proteins” (PR proteins, see, e. g. EP-A 392 225),plant disease resistance genes (e. g. potato cultivars, which expressresistance genes acting against Phytophthora infestans derived from themexican wild potato Solanum bulbocastanum) or T4-lysozym (e. g. potatocultivars capable of synthesizing these proteins with increasedresistance against bacteria such as Erwinia amylvora). The methods forproducing such genetically modified plants are generally known to theperson skilled in the art and are described, e. g. in the publicationsmentioned above.

Furthermore, plants are also covered that are by the use of recombinantDNA techniques capable to synthesize one or more proteins to increasethe productivity (e. g. bio mass production, grain yield, starchcontent, oil content or protein content), tolerance to drought, salinityor other growth-limiting environmental factors or tolerance to pests andfungal, bacterial or viral pathogens of those plants.

Furthermore, plants are also covered that contain by the use ofrecombinant DNA techniques a modified amount of substances of content ornew substances of content, specifically to improve human or animalnutrition, e. g. oil crops that produce health-promoting long-chainomega-3 fatty acids or unsaturated omega-9 fatty acids (e. g. Nexera®rape, DOW Agro Sciences, Canada).

Furthermore, plants are also covered that contain by the use ofrecombinant DNA techniques a modified amount of substances of content ornew substances of content, specifically to improve raw materialproduction, e. g. potatoes that produce increased amounts of amylopectin(e. g. Amflora® potato, BASF SE, Germany).

The mixtures and compositions of the invention, respectively, areparticularly suitable for controlling the following plant diseases:

Albugo spp. (white rust) on ornamentals, vegetables (e. g. A. candida)and sunflowers (e. g. A. tragopogonis); Alternaria spp. (Alternaria leafspot) on vegetables, rape (A. brassicola or brassicae), sugar beets (A.tenuis), fruits, rice, soybeans, potatoes (e. g. A. solani or A.alternata), tomatoes (e. g. A. solani or A. alternata) and wheat;Aphanomyces spp. on sugar beets and vegetables; Ascochyta spp. oncereals and vegetables, e. g. A. tritici (anthracnose) on wheat and A.hordei on barley; Bipolaris and Drechslera spp. (teleomorph:Cochliobolus spp.), e. g. Southern leaf blight (D. maydis) or Northernleaf blight (B. zeicola) on corn, e. g. spot blotch (B. sorokiniana) oncereals and e. g. B. oryzae on rice and turfs; Blumeria (formerlyErysiphe) graminis (powdery mildew) on cereals (e. g. on wheat orbarley); Botrytis cinerea (teleomorph: Botryotinia fuckeliana, greymold) on fruits and berries (e. g. strawberries), vegetables (e. g.lettuce, carrots, celery and cabbages), rape, flowers, vines, forestryplants and wheat; Bremia lactucae (downy mildew) on lettuce;Ceratocystis (syn. Ophiostoma) spp. (rot or wilt) on broad-leaved treesand evergreens, e. g. C. ulmi (Dutch elm disease) on elms; Cercosporaspp. (Cercospora leaf spots) on corn (e. g. Gray leaf spot: C.zeae-maydis), rice, sugar beets (e. g. C. beticola), sugar cane,vegetables, coffee, soybeans (e. g. C. sojina or C. kikuchii) and rice;Cladosporium spp. on tomatoes (e. g. C. fulvum: leaf mold) and cereals,e. g. C. herbarum (black ear) on wheat; Claviceps purpurea (ergot) oncereals; Cochliobolus (anamorph: Helminthosporium of Bipolaris) spp.(leaf spots) on corn (C. carbonum), cereals (e. g. C. sativus, anamorph:B. sorokiniana) and rice (e. g. C. miyabeanus, anamorph: H. oryzae);Colletotrichum (teleomorph: Glomerella) spp. (anthracnose) on cotton (e.g. C. gossypii), corn (e. g. C. graminicola: Anthracnose stalk rot),soft fruits, potatoes (e. g. C. coccodes: black dot), beans (e. g. C.lindemuthianum) and soybeans (e. g. C. truncatum or C. gloeosporioides);Corticium spp., e. g. C. sasakii (sheath blight) on rice; Corynesporacassilicola (leaf spots) on soybeans and ornamentals; Cycloconium spp.,e. g. C. oleaginum on olive trees; Cylindrocarpon spp. (e. g. fruit treecanker or young vine decline, teleomorph: Nectria or Neonectria spp.) onfruit trees, vines (e. g. C. lirodendri, teleomorph: Neonectrialiriodendrii: Black Foot Disease) and ornamentals; Dematophora(teleomorph: Rosellinia) necatrix (root and stem rot) on soybeans;Diaporthe spp., e. g. D. phaseolorum (damping off) on soybeans;Drechslera (syn. Helminthosporium, teleomorph: Pyrenophora) spp. oncorn, cereals, such as barley (e. g. D. teres, net blotch) and wheat (e.g. D. tritici-repentis: tan spot), rice and turf; Esca (dieback,apoplexy) on vines, caused by Formitiporia (syn. Phellinus) punctata, F.mediterranea, Phaeomoniella chlamydospora (earlier Phaeoacremoniumchlamydosporum), Phaeoacremonium aleophilum and/or Botryosphaeriaobtusa; Elsinoe spp. on pome fruits (E. pyri), soft fruits (E. veneta:anthracnose) and vines (E. ampelina: anthracnose); Entyloma oryzae (leafsmut) on rice; Epicoccum spp. (black mold) on wheat; Erysiphe spp.(powdery mildew) on sugar beets (E. betae), vegetables (e. g. E. pisi),such as cucurbits (e. g. E. cichoracearum), cabbages, rape (e. g. E.cruciferarum); Eutypa lata (Eutypa canker or dieback, anamorph:Cytosporina lata, syn. Libertella blepharis) on fruit trees, vines andornamental woods; Exserohilum (syn. Helminthosporium) spp. on corn (e.g. E. turcicum); Fusarium (teleomorph: Gibberella) spp. (wilt, root orstem rot) on various plants, such as F. graminearum or F. culmorum (rootrot, scab or head blight) on cereals (e. g. wheat or barley), F.oxysporum on tomatoes, F. solani (f. sp. glycines now syn. F.virguliforme) and F. tucumaniae and F. brasiliense each causing suddendeath syndrome on soybeans, and F. verticillioides on corn;Gaeumannomyces graminis (take-all) on cereals (e. g. wheat or barley)and corn; Gibberella spp. on cereals (e. g. G. zeae) and rice (e. g. G.fujikuroi: Bakanae disease); Glomerella cingulata on vines, pome fruitsand other plants and G. gossypii on cotton; Grainstaining complex onrice; Guignardia bidwellii (black rot) on vines; Gymnosporangium spp. onrosaceous plants and junipers, e. g. G. sabinae (rust) on pears;Helminthosporium spp. (syn. Drechslera, teleomorph: Cochliobolus) oncorn, cereals and rice; Hemileia spp., e. g. H. vastatrix (coffee leafrust) on coffee; Isariopsis clavispora (syn. Cladosporium vitis) onvines; Macrophomina phaseolina (syn. phaseoli) (root and stem rot) onsoybeans and cotton; Microdochium (syn. Fusarium) nivale (pink snowmold) on cereals (e. g. wheat or barley); Microsphaera diffusa (powderymildew) on soybeans; Monilinia spp., e. g. M. taxa, M. fructicola and M.fructigena (bloom and twig blight, brown rot) on stone fruits and otherrosaceous plants; Mycosphaerella spp. on cereals, bananas, soft fruitsand ground nuts, such as e. g. M. graminicola (anamorph: Septoriatritici, Septoria blotch) on wheat or M. fijiensis (black Sigatokadisease) on bananas; Peronospora spp. (downy mildew) on cabbage (e. g.P. brassicae), rape (e. g. P. parasitica), onions (e. g. P. destructor),tobacco (P. tabacina) and soybeans (e. g. P. manshurica); Phakopsorapachyrhizi and P. meibomiae (soybean rust) on soybeans; Phialophora spp.e. g. on vines (e. g. P. tracheiphila and P. tetraspora) and soybeans(e. g. P. gregata: stem rot); Phoma lingam (root and stem rot) on rapeand cabbage and P. betae (root rot, leaf spot and damping-off) on sugarbeets; Phomopsis spp. on sunflowers, vines (e. g. P. viticola: can andleaf spot) and soybeans (e. g. stem rot: P. phaseoli, teleomorph:Diaporthe phaseolorum); Physoderma maydis (brown spots) on corn;Phytophthora spp. (wilt, root, leaf, fruit and stem root) on variousplants, such as paprika and cucurbits (e. g. P. capsici), soybeans (e.g. P. megasperma, syn. P. sojae), potatoes and tomatoes (e. g. P.infestans late blight) and broad-leaved trees (e. g. P. ramorum: suddenoak death); Plasmodiophora brassicae (club root) on cabbage, rape,radish and other plants; Plasmopara spp., e. g. P. viticola (grapevinedowny mildew) on vines and P. halstedii on sunflowers; Podosphaera spp.(powdery mildew) on rosaceous plants, hop, pome and soft fruits, e. g.P. leucotricha on apples; Polymyxa spp., e. g. on cereals, such asbarley and wheat (P. graminis) and sugar beets (P. betae) and therebytransmitted viral diseases; Pseudocercosporella herpotrichoides(eyespot, teleomorph: Tapesia yallundae) on cereals, e. g. wheat orbarley; Pseudoperonospora (downy mildew) on various plants, e. g. P.cubensis on cucurbits or P. humili on hop; Pseudopezicula tracheiphilia(red fire disease or ‘rotbrenner’, anamorph: Phialophora) on vines;Puccinia spp. (rusts) on various plants, e. g. P. triticina (brown orleaf rust), P. striiformis (stripe or yellow rust), P. hordei (dwarfrust), P. graminis (stem or black rust) or P. recondita (brown or leafrust) on cereals, such as e. g. wheat, barley or rye, P. kuehnii (orangerust) on sugar cane and P. asparagi on asparagus; Pyrenophora (anamorph:Drechslera) tritici-repentis (tan spot) on wheat or P. teres (netblotch) on barley; Pyricularia spp., e. g. P. olyzae (teleomorph:Magnaporthe grisea, rice blast) on rice and P. grisea on turf andcereals; Pythium spp. (damping-off) on turf, rice, corn, wheat, cotton,rape, sunflowers, soybeans, sugar beets, vegetables and various otherplants (e. g. P. ultimum or P. aphanidermatum); Ramularia spp., e. g. R.collo-cygni (Rannularia leaf spots, Physiological leaf spots) on barleyand R. beticola on sugar beets; Rhizoctonia spp. on cotton, rice,potatoes, turf, corn, rape, potatoes, sugar beets, vegetables andvarious other plants, e. g. R. solani (root and stem rot) on soybeans,R. solani (sheath blight) on rice or R. cerealis (Rhizoctonia springblight) on wheat or barley; Rhizopus stolonifer (black mold, soft rot)on strawberries, carrots, cabbage, vines and tomatoes; Rhynchosporiumsecalis (scald) on barley, rye and triticale; Sarocladium oryzae and S.attenuatum (sheath rot) on rice; Sclerotinia spp. (stem rot or whitemold) on vegetables and field crops, such as rape, sunflowers (e. g. S.sclerotiorum) and soybeans (e. g. S. rolfsii or S. sclerotiorum);Septoria spp. on various plants, e. g. S. glycines (brown spot) onsoybeans, S. tritici (Septoria blotch) on wheat and S. (syn.Stagonospora) nodorum (Stagonospora blotch) on cereals; Uncinula (syn.Erysiphe) necator(powdery mildew, anamorph: Oidium tuckeri) on vines;Setospaeria spp. (leaf blight) on corn (e. g. S. turcicum, syn.Helminthosporium turcicum) and turf; Sphacelotheca spp. (smut) on corn,(e. g. S. reiliana: head smut), sorghum and sugar cane; Sphaerothecafuliginea (powdery mildew) on cucurbits; Spongospora subterranea(powdery scab) on potatoes and thereby transmitted viral diseases;Stagonospora spp. on cereals, e. g. S. nodorum (Stagonospora blotch,teleomorph: Leptosphaeria [syn. Phaeosphaeria]; nodorum) on wheat;Synchytrium endobioticum on potatoes (potato wart disease); Taphrinaspp., e. g. T. deformans (leaf curl disease) on peaches and T. pruni(plum pocket) on plums; Thielaviopsis spp. (black root rot) on tobacco,pome fruits, vegetables, soybeans and cotton, e. g. T. basicola (syn.Chalara elegans); Tilletia spp. (common bunt or stinking smut) oncereals, such as e. g. T. tritici (syn. T. caries, wheat bunt) and T.controversa (dwarf bunt) on wheat; Typhula incamata (grey snow mold) onbarley or wheat; Urocystis spp., e. g. U. occulta (stem smut) on rye;Uromyces spp. (rust) on vegetables, such as beans (e. g. U.appendiculatus, syn. U. phaseoli) and sugar beets (e. g. U. betae);Ustilago spp. (loose smut) on cereals (e. g. U. nuda and U. avaenae),corn (e. g. U. maydis: corn smut) and sugar cane; Venturia spp. (scab)on apples (e. g. V. inaequalis) and pears; and Verticillium spp. (wilt)on various plants, such as fruits and ornamentals, vines, soft fruits,vegetables and field crops, e. g. V. dahliae on strawberries, rape,potatoes and tomatoes.

The mixtures and compositions of the invention, respectively, are alsosuitable for controlling harmful pathogens, especially fungi, in theprotection of stored products or harvest and in the protection ofmaterials. The term “protection of materials” is to be understood todenote the protection of technical and non-living materials, such asadhesives, glues, wood, paper and paperboard, textiles, leather, paintdispersions, plastics, cooling lubricants, fiber or fabrics, against theinfestation and destruction by harmful microorganisms, such as fungi andbacteria. As to the protection of wood and other materials, theparticular attention is paid to the following harmful fungi: Ascomycetessuch as Ophiostoma spp., Ceratocystis spp., Aureobasidium pullulans,Sclerophoma spp., Chaetomium spp., Humicola spp., Petriella spp.,Trichurus spp.; Basidiomycetes such as Coniophora spp., Coriolus spp.,Gloeophyllum spp., Lentinus spp., Pleurotus spp., Poria spp., Serpulaspp. and Tyromyces spp., Deuteromycetes such as Aspergillus spp.,Cladosporium spp., Penicillium spp., Trichorma spp., Alternaria spp.,Paecllomyces spp. and Zygomycetes such as Mucor spp., and in addition inthe protection of stored products and harvest the following yeast fungiare worthy of note: Candida spp. and Saccharomyces cerevisiae. Themethod of treatment according to the invention can also be used in thefield of protecting stored products or harvest against attack of fungiand microorganisms. According to the present invention, the term “storedproducts” is understood to denote natural substances of plant or animalorigin and their processed forms, which have been taken from the naturallife cycle and for which long-term protection is desired. Storedproducts of crop plant origin, such as plants or parts thereof, forexample stalks, leafs, tubers, seeds, fruits or grains, can be protectedin the freshly harvested state or in processed form, such as pre-dried,moistened, comminuted, ground, pressed or roasted, which process is alsoknown as post-harvest treatment. Also falling under the definition ofstored products is timber, whether in the form of crude timber, such asconstruction timber, electricity pylons and barriers, or in the form offinished articles, such as furniture or objects made from wood. Storedproducts of animal origin are hides, leather, furs, hairs and the like.The combinations according the present invention can preventdisadvantageous effects such as decay, discoloration or mold. Preferably“stored products” is understood to denote natural substances of plantorigin and their processed forms, more preferably fruits and theirprocessed forms, such as pomes, stone fruits, soft fruits and citrusfruits and their processed forms.

The mixtures and compositions according to the invention areparticularly important in the control of a multitude of phytopathogenicinsects or other pests (e.g. lepidopterans, beetles, dipterans, thrips,heteropterans, hemiptera, homoptera, termites, orthopterans, arachnids,and nematodes) on various cultivated plants, such as cereals, e. g.wheat, rye, barley, triticale, oats or rice; beet, e. g. sugar beet orfodder beet; fruits, such as pomes, stone fruits or soft fruits, e. g.apples, pears, plums, peaches, almonds, cherries, strawberries,raspberries, blackberries or gooseberries; leguminous plants, such aslentils, peas, alfalfa or soybeans; oil plants, such as rape, mustard,olives, sunflowers, coconut, cocoa beans, castor oil plants, oil palms,ground nuts or soybeans; cucurbits, such as squashes, cucumber ormelons; fiber plants, such as cotton, flax, hemp or jute; citrus fruit,such as oranges, lemons, grapefruits or mandarins; vegetables, such asspinach, lettuce, asparagus, cabbages, carrots, onions, tomatoes,potatoes, cucurbits or paprika; lauraceous plants, such as avocados,cinnamon or camphor; energy and raw material plants, such as corn,soybean, rape, sugar cane or oil palm; corn; tobacco; nuts; coffee; tea;bananas; vines (table grapes and grape juice grape vines); hop; turf;natural rubber plants or ornamental and forestry plants, such asflowers, shrubs, broad-leaved trees or evergreens, e. g. conifers; andon the plant propagation material, such as seeds, and the crop materialof these plants.

Preferably the inventive mixtures and compositions are used forcontrolling a multitude of pests on field crops, such as potatoes sugarbeets, tobacco, wheat, rye, barley, oats, rice, corn, cotton, soybeans,rape, legumes, sunflowers, coffee or sugar cane; fruits; vines;ornamentals; or vegetables, such as cucumbers, tomatoes, beans orsquashes.

The inventive mixtures and the compositions thereof, respectively, areparticularly suitable for controlling the following harmful insects fromthe order of the

lepidopterans (Lepidoptera), for example AgrotiS ypshon, AgrotiSsegetum, Alabama argillacea, Anticarsia gemmatalis, Argyresthiaconjugella, Autographa gamma, Bupalus piniarius, Cacoecia murinana,Capua reticulana, Cheimatobia brumata, Choristoneura fumiferana,Choristoneura occidentalis, Cirphis unipuncta, Cydia pomonella,Dendrolimus pini, Diaphania nitidalis, Diatraea grandiosella, Eariasinsulana, Elasmopalpus lignosellus, Eupoechia ambiguella, Evetriabouliana, Feltia subterranea, Galleria mellonella, Grapholithafunebrana, Grapholitha molesta, Heliothis armigera, Heliothis virescens,Heliothis zea, Hellula undalis, Hibernia defoliaria, Hyphantria cunea,Hyponomeuta malinellus, Keiferia lycopersicella, Lambdina fiscellaria,Laphygma exigua, Leucoptera coffeella, Leucoptera scitella,Lithocolletis blancardella, Lobesia botrana, Loxostege sticticalis,Lymantria dispar, Lymantria monacha, Lyonetia clerkella, Malacosomaneustria, Mamestra brassicae, Orgyia pseudotsugata, Ostrinia nubilalis,Panolis flammea, Pectinophora gossypiella, Peridroma saucia, Phalerabucephala, Phthorimaea operculella, Phyllocnistis citrella, Pierisbrassicae, Plathypena scabra, Plutella xylostella, Pseudoplusiaincludens, Rhyacionia frustrana, Scrobipalpula absoluta, Sitotrogacerealella, SparganothIS pheriana, Spodoptera frugiperda, Spodopteralittoralis, Spodoptera litura, Thaumatopoea pityocampa, Tortrixviridana, Trichoplusia ni and Zeiraphera canadensis,

beetles (Coleoptera), for example Agrilus sinuatus, Agriotes lineatus,Agriotes obscurus, Amphimallus solstitialis, Anisandrus dispar,Anthonomus grandis, Anthonomus pomorum, Atomaria linearis, Blastophaguspiniperda, Blitophaga undata, Bruchus rufimanus, Bruchus pisorum,Bruchus lentis, Byctiscus betulae, Cassida nebulosa, Cerotomatrifurcata, Ceuthorrhynchus assimilis, Ceuthorrhynchus napi, Chaetocnematibialis, Conoderus vespertinus, Criocen:s asparagi, Diabroticalongicornis, Diabrotica speciosa, Diabrotica 12-punctata, Diabroticavirgifera, Diloboderus abderus, Ephachna varivestis, Epitrixhirtipennis, Eutinobothrus brasiliensis, Hylobius abietis, Hyperabrunneipennis, Hypera postica, Ips typographus, Lema bilineata, Lemamelanopus, Leptinotarsa decemlineata, Limonius californicus,Lissorhoptrus olyzophhus, Melanotus communis, Meligethes aeneus,Melolontha hippocastani, Melolontha melolontha, Oulema oryzae,Ortiorrhynchus sulcatus, Oryazophagus oryzae, Otiorrhynchus ovatus,Phaedon cochleariae, Phyllotreta chrysocephala, Phyllophaga sp.,Phyllophaga cuyabana, Phyllophaga triticophaga, Phyllopertha horticola,Phyllotreta nemorum, Phyllotreta striolata, Popilia japonica, Sitonalineatus and Sitophllus granaria,

dipterans (Diptera), for example Aedes aegypti, Aedes vexans, Anastrephaludens, Anopheles maculipennis, Ceratitis capitata, Chrysomya bezziana,Chrysomya hominivorax, Chrysomya macellaria, Contarinia sorghicola,Cordylobia anthropophaga, Culex pipiens, Dacus cucurbitae, Dacus oleae,Dasineura brassicae, Fannia canicularis, Gasterophilus intestinalis,Glossina morsitans, Haematobia irritans, Haplodiplosid equestris,Hylemyia platura, Hypoderma lineata, Liriomyza sativae, Liriomyzatrifolil, Lucilia caprina, Lucllia cuprina, Lucllia sericata, Lycoriapectoralis, Mayetiola destructor, Musca domestica, Muscina stabulans,Oestrus ovis, Oscinella frit, Pegomya hysocyami, Phorbia antiqua,Phorbia brassicae, Phorbia coarctata, Rhagoietis cerasi, Rhagoletispomonella, Tabanus bovinus, Tipula oleracea and Tipula paludosa,

thrips (Thysanoptera), e.g. Frankliniella fusca, Frankliniellaoccidentalis, Frankliniella tritici, Scirtothrips citri, Thrips oryzae,Thrips palmi and Thrips tabaci,

hymenopterans (Hymenoptera), e.g. Acromyrmex ambuguus, Acromyrmexcrassispinus, Acromyrmex heiery, Acromyrmex landolti, Acromyrmexsubterraneus, Athalia rosae, Atta capiguara, Atta cephalotes, Attalaevigata, Atta robusta, Atta sexdens, Atta texana, Hoplocampa minuta,Hoplocampa testudinea, Monomorium Pharaonis, Solenopsis geminata andSolenopsis invicta,

heteropterans (Heteroptera), e.g. Acrosternum hilre, Blissusleucopterus, Cyrtopeltis notatus, Dichelops furcatus, Dysdercuscingulatus, Dysdercus intermedius, Euchistos heros, Eurygasterintegriceps, Euschistus impictiventris, Leptoglossus phyllopus, Lyguslineolaris, Lygus pratensis, Nezara viridula, Piesma quadrata,Piezodorus guildini, Solubea insularis and Thyanta perditor,

Hemiptera and Homoptera, e.g. Acrosternum hilre, Blissus leucopterus,Cyrtopeltis notatus, Diaphorina citri, Dysdercus cingulatus, Dysdercusintermedius, Eurygaster integriceps, Euschistus impictiventris,Leptoglossus phyllopus, Lygus lineolaris, Lygus pratensis, Nezaraviridula, Piesma quadrata, Solubea insularis, Thyanta perditor,Acyrthosiphon onobrychis, Adelges laricis, Aphidula nasturtii, Aphisfabae, Aphis forbesi, Aphis pony, Aphis gossypi, Aphis grossulariae,Aphis schneiden, Aphis spiraecola, Aphis sambuci, Acyrthosiphon pisum,Aulacorthum solani, Brachycaudus cardui, Brachycaudus hekhlysi,Brachycaudus persicae, Brachycaudus Arunicola, Brevicoryne brassicae,Capitophorus horni, Cerosipha gossypii, Chaetosiphon fragaefolii,Cryptomyzus ribis, Dreyfusia nordmannianae, Dreyfusia piceae, Dysaphisradicola, Dysaulacorthum pseudosolani, Dysaphis plantaginea, Dysaphisprti, Empoasca fabae, Hyalopterus pruni, Hyperomyzus lactucae,Macrosiphum avenae, Macrosiphum euphorbiae, Macrosiphon rosae, Megouraviciae, Melanaphis pyrarius, Metopolophium dirhodum, Myzodes persicae,Myzus ascalonicus, Myzus cerasi, Myzus varians, Nasonovia ribis-nigri,Nilaparvata lugens, Pemphigus bursarius, Perkinsiella saccharicida,Phorodon humuli, Psylla mali, Psylla piri, Rhopalomyzus ascalonicus,Rhopalosiphum maidis, Rhopalosiphum padi, Rhopalosiphum insertum,Sappaphis mala, Sappaphis mall, Schizaphis graminum, Schizoneuralanuginosa, Sitobion avenae, Trialeurodes vaporariorum, Toxopteraaurantiiand, Viteus vitifolil, Cimex lectularius, Cimex hemipterus,Reduvius senilis, Triatoma spp., and Arilus critatus,

termites (Isoptera), e.g. Calotermes flavicollis, Cornitermes cumulans,Heterotermes tenuis, Leucotermes flavipes, Neocapritemes opacus,Procornitermes triacifer; Reticulitermes lucifugus, Syntermes molestus,and Termes natalensis,

orthopterans (Orthoptera), e.g. Acheta domestica, Blatta orientalis,Blattella germanica, Forticula auricularia, Gryllotalpa gryllotalpa,Locusta migratoria, Melanoplus bivittatus, Melanoplus femur-rubrum,Melanoplus mexicanus, Melanoplus sanguinipes, Melanoplus spretus,Nomadacris septemfasciata, Periplaneta americana, Schistocercaamericana, Schistocerca peregrina, Stauronotus maroccanus and Tachycinesasynamorus,

Arachnoidea, such as arachnids, e.g. of the families Argasidae, Ixodidaeand Sarcoptidae, such as Amblyomma americanum, Amblyomma variegatum,Argas persicus, Boophilus annulatus, Boophilus decoloratus, Boophilusmicroplus, Dermacentor silvarum, Hyalomma truncatum, Ixodes ricinus,Ixodes rubicundus, Ornithodorus moubata, Otobius megnini, Dermanyssusgallinae, Psoroptes ovis, Rhipicephalus appendiculatus, Rhipicephalusevertsi, Sarcoptes scabiei, and Eriophyidae spp. such as Aculusschlechtendali, Phyllocoptrata oleivora and Eriophyes sheldoni;Tarsonemidae spp. such as Phytonemus pallidus and Polyphagotarsonemuslatus; Tenuipalpidae spp. such as Brevipalpus phoenicis; Tetranychidaespp. such as Tetranychus cinnabarinus, Tetranychus kanzawai, Tetranychuspacificus, Tetranychus telarius and Tetranychus urticae, Panonychusulmi, Panonychus citri, and Oligonychus pratensis.

In particular, the inventive mixtures are suitable for combating pestsof the orders Coleoptera, Lepidoptera, Thysanoptera, Homoptera,Isoptera, and Orthoptera.

They are also suitable for controlling the following plant parasiticnematodes such as root-knot nematodes, Meloidogyne arenaria, Meloidogynechitwoodi, Meloidogyne exigua, Meloidogyne hapla, Meloidogyne incognita,Meloidogyne javanica and other Meloidogyne species; cyst nematodes,Globodera rostochiensis, Globodera pallida, Globodera tabacum and otherGlobodera species, Heterodera avenae, Heterodera glycines, Heteroderaschachtii, Heterodera trifolii, and other Heterodera species; seed gallnematodes, Anguina funesta, Anguina tritici and other Anguina species;stem and foliar nematodes, Aphelenchoides besseyi, Aphelenchoidesfragariae, Aphelenchoides ritzemabosi and other Aphelenchoides species;sting nematodes, Belonolaimus longicaudatus and other Belonolaimusspecies; pine nematodes, Bursapheienchus xylophilus and otherBursaphelenchus species; ring nematodes, Criconema species, Criconemellaspecies, Criconemoides species, and Mesocriconema species; stem and bulbnematodes, Dityienchus destructor, Dityienchus dipsaci, Dityienchusmyceliophagus and other Dityienchus species; awl nematodes, Dolichodorusspecies; spiral nematodes, Helicotylenchus dihystera, Helicotylenchusmulticinctus and other Helicotylenchus species, Rotylenchus robustus andother Rotylenchus species; sheath nematodes, Hemicycliophora species andHemicriconemoides species; Hirshmanniella species; lance nematodes,Hoplolaimus columbus, Hoplolaimus galeatus and other Hoplolaimusspecies; false root-knot nematodes, Nacobbus aberrans and other Nacobbusspecies; needle nematodes, Longidorus elongates and other Longidorusspecies, pin nematodes, Paratylenchus species; lesion nematodes,Pratylenchus brachyurus, Pratylenchus coffeae, Pratylenchus curvitatus,Pratylenchus goodeyi, Pratylencus neglectus, Pratylenchus penetrans,Pratylenchus scribneri, Pratylenchus vulnus, Pratylenchus zeae and otherPratylenchus species, Radinaphelenchus cocophilus and otherRadinaphelenchus species, burrowing nematodes, Radopholus similis andother Radopholus species, reniform nematodes, Rotylenchulus reniformisand other Rotylenchulus species, Scutellonema species, stubby rootnematodes, Trichodorus primitivus and other Trichodorus species,Paratrichodorus minor and other Paratrichodorus species, stuntnematodes, Tylenchorhynchus claytoni, Tylenchorhynchus dubius and otherTylenchorhynchus species and Merlinius species; citrus nematodes,Tylenchulus semipenetrans and other Tylenchulus species, daggernematodes, Xiphinema americanum, Xiphinema index, Xiphinemadiversicaudatum and other Xiphinema species; and other plant parasiticnematode species

In an equally preferred embodiment, the present invention relates to amethod for controlling animal pests (insects, acarids or nematodes),wherein the animal pests (insects, acarids or nematodes), their habitat,breeding grounds, their locus or the plants to be protected againstanimal pest (insects, acarids or nematodes) attack are treated with aneffective amount of an inventive mixture comprising a Paenibacillusstrain as defined above and biopesticide II.

In general, “pesticidally effective amount” means the amount of theinventive mixtures or of compositions comprising the mixtures needed toachieve an observable effect on growth, including the effects ofnecrosis, death, retardation, prevention, and removal, destruction, orotherwise diminishing the occurrence and activity of the targetorganism. The pesticidally effective amount can vary for the variousmixtures/compositions used in the invention. A pesticidally effectiveamount of the mixtures/compositions will also vary according to theprevailing conditions such as desired pesticidal effect and duration,weather, target species, locus, mode of application, and the like.

Plant propagation materials may be treated with the mixtures andcompositions of the invention prophylactically either at or beforeplanting or transplanting.

In particular, the present invention relates to a method for protectionof plant propagation material from pests, wherein the plant propagationmaterial is treated with an effective amount of an inventive mixture.

In an equally preferred embodiment, the present invention relates to amethod for protection of plant propagation material from harmful fungi,wherein the plant propagation material is treated with an effectiveamount of an inventive mixture.

In an equally preferred embodiment, the present invention relates to amethod for improving the health of plants, wherein the plants aretreated with an effective amount of an inventive mixture.

The term “plant health effective amount” denotes an amount of theinventive mixtures, which is sufficient for achieving plant healtheffects as defined herein below. More exemplary information aboutamounts, ways of application and suitable ratios to be used is givenbelow. Anyway, the skilled artisan is well aware of the fact that suchan amount can vary in a broad range and is dependent on various factors,e.g. the treated cultivated plant or material and the climaticconditions.

Healthier plants are desirable since they result among others in betteryields and/or a better quality of the plants or crops, specificallybetter quality of the harvested plant parts. Healthier plants alsobetter resist to biotic and/or abiotic stress. A high resistance againstbiotic stresses in turn allows the person skilled in the art to reducethe quantity of pesticides applied and con-consequently to slow down thedevelopment of resistances against the respective pesticides.

It was therefore an object of the present invention to provide apesticidal composition which solves the problems outlined above, andwhich should, in particular, improve the health of plants, in particularthe yield of plants.

The term “health of a plant” or “plant health” is defined as a conditionof the plant and/or its products which is determined by several aspectsalone or in combination with each other such as increased yield, plantvigor, quality of harvested plant parts and tolerance to abiotic and/orbiotic stress.

It has to be emphasized that the above mentioned effects of theinventive mixtures, i.e. enhanced health of the plant, are also presentwhen the plant is not under biotic stress and in particular when theplant is not under pest pressure.

For seed treatment e.g. as inoculant and/or foliar application forms, itis evident that a plant suffering from fungal or insecticidal attackproduces a smaller biomass and leads to a reduced yield as compared to aplant which has been subjected to curative or preventive treatmentagainst the pathogenic fungus or any other relevant pest and which cangrow without the damage caused by the biotic stress factor. However, themethods according to the invention lead to an enhanced plant health evenin the absence of any biotic stress. This means that the positiveeffects of the mixtures of the invention cannot be explained just by thepesticidal activities of the bacterial strains of component 1) and thebiopesticide II, but are based on further activity profiles.Accordingly, the application of the inventive mixtures can also becarried out in the absence of pest pressure.

Each plant health indicator listed below, which is selected from thegroups consisting of yield, plant vigor, quality and tolerance of theplant to abiotic and/or biotic stress, is to be understood as apreferred embodiment of the present invention either each on its own orpreferably in combination with each other.

According to the present invention, “increased yield” of a plant meansthat the yield of a product of the respective plant is increased by ameasurable amount over the yield of the same product of the plantproduced under the same conditions, but without the application of theinventive mixture.

For seed treatment e.g. as inoculant and/or foliar application forms,increased yield can be characterized, among others, by the followingimproved properties of the plant: increased plant weight; and/orincreased plant height; and/or increased biomass such as higher overallfresh weight (FW); and/or increased number of flowers per plant; and/orhigher grain and/or fruit yield; and/or more tillers or side shoots(branches); and/or larger leaves; and/or increased shoot growth; and/orincreased protein content; and/or increased oil content; and/orincreased starch content; and/or increased pigment content; and/orincreased chlorophyll content (chlorophyll content has a positivecorrelation with the plant's photosynthesis rate and accordingly, thehigher the chlorophyll content the higher the yield of a plant) and/orincreased quality of a plant.

“Grain” and “fruit” are to be understood as any plant product which isfurther utilized after harvesting, e.g. fruits in the proper sense,vegetables, nuts, grains, seeds, wood (e.g. in the case of silvicultureplants), flowers (e.g. in the case of gardening plants, ornamentals)etc., that is anything of economic value that is produced by the plant.

According to the present invention, the yield is increased by at least4%. In general, the yield increase may even be higher, for example 5 to10%, more preferable by 10 to 20%, or even 20 to 30%

According to the present invention, the yield—if measured in the absenceof pest pressure—is increased by at least 2% In general, the yieldincrease may even be higher, for example until 4% to 5% or even more.

Another indicator for the condition of the plant is the plant vigor. Theplant vigor becomes manifest in several aspects such as the generalvisual appearance.

For foliar applications, improved plant vigor can be characterized,among others, by the following improved properties of the plant:improved vitality of the plant; and/or improved plant growth; and/orimproved plant development; and/or improved visual appearance; and/orimproved plant stand (less plant verse/lodging and/or bigger leaf blade;and/or bigger size; and/or increased plant height; and/or increasedtiller number; and/or increased number of side shoots; and/or increasednumber of flowers per plant; and/or increased shoot growth; and/orenhanced photosynthetic activity (e.g. based on increased stomatalconductance and/or increased CO2 assimilation rate)); and/or earlierflowering; and/or earlier fruiting; and/or earlier grain maturity;and/or less non-productive tillers; and/or less dead basal leaves;and/or less input needed (such as fertilizers or water); and/or greenerleaves; and/or complete maturation under shortened vegetation periods;and/or easier harvesting; and/or faster and more uniform ripening;and/or longer shelf-life; and/or longer panicles; and/or delay ofsenescence; and/or stronger and/or more productive tillers; and/orbetter extractability of ingredients; and/or improved quality of seeds(for being seeded in the following seasons for seed production); and/orreduced production of ethylene and/or the inhibition of its reception bythe plant.

Another indicator for the condition of the plant is the “quality” of aplant and/or its products. According to the present invention, enhancedquality means that certain plant characteristics such as the content orcomposition of certain ingredients are increased or improved by ameasurable or noticeable amount over the same factor of the plantproduced under the same conditions, but without the application of themixtures of the present invention. Enhanced quality can becharacterized, among others, by following improved properties of theplant or its product: increased nutrient content; and/or increasedprotein content; and/or increased oil content; and/or increased starchcontent; and/or increased content of fatty acids; and/or increasedmetabolite content; and/or increased carotenoid content; and/orincreased sugar content; and/or increased amount of essential aminoacids; and/or improved nutrient composition; and/or improved proteincomposition; and/or improved composition of fatty acids; and/or improvedmetabolite composition; and/or improved carotenoid composition; and/orimproved sugar composition; and/or improved amino acids composition;and/or improved or optimal fruit color; and/or improved leaf color;and/or higher storage capacity; and/or better processability of theharvested products.

Another indicator for the condition of the plant is the plants toleranceor resistance to biotic and/or abiotic stress factors. Biotic andabiotic stress, especially over longer terms, can have harmful effectson plants.

Biotic stress is caused by living organisms while abiotic stress iscaused for example by environmental extremes. According to the presentinvention, “enhanced tolerance or resistance to biotic and/or abioticstress factors” means (1.) that certain negative factors caused bybiotic and/or abiotic stress are diminished in a measurable ornoticeable amount as compared to plants exposed to the same conditions,but without being treated with an inventive mixture and (2.) that thenegative effects are not diminished by a direct action of the inventivemixture on the stress factors, e.g. by its fungicidal or insecticidalaction which directly destroys the microorganisms or pests, but ratherby a stimulation of the plants' own defensive reactions against saidstress factors.

Negative factors caused by biotic stress such as pathogens and pests arewidely known and are caused by living organisms, such as competingplants (for example weeds), microorganisms (such as phythopathogenicfungi and/or bacteria) and/or viruses.

Negative factors caused by abiotic stress are also well-known and canoften be observed as reduced plant vigor (see above), for example:

less yield and/or less vigor, for both effects examples can be burnedleaves, less flowers, pre-mature ripening, later crop maturity, reducednutritional value amongst others.

Abiotic stress can be caused for example by: extremes in temperaturesuch as heat or cold (heat stress/cold stress); and/or strong variationsin temperature; and/or temperatures unusual for the specific season;and/or drought (drought stress); and/or extreme wetness; and/or highsalinity (salt stress); and/or radiation (for example by increased UVradiation due to the decreasing ozone layer); and/or increased ozonelevels (ozone stress); and/or organic pollution (for example byphythotoxic amounts of pesticides); and/or inorganic pollution (forexample by heavy metal contaminants).

As a result of biotic and/or abiotic stress factors, the quantity andthe quality of the stressed plants decrease. As far as quality (asdefined above) is concerned, reproductive development is usuallyseverely affected with consequences on the crops which are important forfruits or seeds. Synthesis, accumulation and storage of proteins aremostly affected by temperature; growth is slowed by almost all types ofstress; polysaccharide synthesis, both structural and storage is reducedor modified: these effects result in a decrease in biomass (yield) andin changes in the nutritional value of the product.

As pointed out above, the above identified indicators for the healthcondition of a plant may be interdependent and may result from eachother. For example, an increased resistance to biotic and/or abioticstress may lead to a better plant vigor, e.g. to better and biggercrops, and thus to an increased yield. Inversely, a more developed rootsystem may result in an increased resistance to biotic and/or abioticstress. However, these interdependencies and interactions are neitherall known nor fully understood and therefore the different indicatorsare described separately.

In one embodiment the inventive mixtures effectuate an increased yieldof a plant or its product. In another embodiment the inventive mixtureseffectuate an increased vigor of a plant or its product. In anotherembodiment the inventive mixtures effectuate in an increased quality ofa plant or its product. In yet another embodiment the inventive mixtureseffectuate an increased tolerance and/or resistance of a plant or itsproduct against biotic stress. In yet another embodiment the inventivemixtures effectuate an increased tolerance and/or resistance of a plantor its product against abiotic stress.

The invention also relates to agrochemical compositions comprising anauxiliary and at least one Paenibacillus strain as defined herein, or acell-free extract thereof or at least one metabo-metabolite thereof, andat least one biopesticide II according to the invention.

An agrochemical composition comprises a fungicidally or insecticidallyeffective amount of at least one Paenibacillus strain as defined herein,or a cell-free extract thereof or at least one metabolite thereof, andat least one biopesticide II. The term “effective amount” denotes anamount of the composition or of at least one Paenibacillus strain asdefined herein, or a cell-free extract thereof or at least onemetabolite thereof, and at least one biopesticide II, which issufficient for promoting plant health, controlling harmful fungi orharmful pests on cultivated plants or in the protection of materials andwhich does not result in a substantial damage to the treated plants ormaterials. Such an amount can vary in a broad range and is dependent onvarious factors, such as the fungal or pest species to be controlled,the treated cultivated plant or material, the climatic conditions.

The at least one Paenibacillus strain as defined herein, or a cell-freeextract thereof or at least one metabolite thereof, and at least onebiopesticide II can be converted into customary types of agrochemicalcompositions, e. g. solutions, emulsions, suspensions, dusts, powders,pastes, granules, pressings, capsules, and mixtures thereof. Examplesfor composition types are suspensions (e.g. SC, OD, FS), emulsifiableconcentrates (e.g. EC), emulsions (e.g. EW, EO, ES, ME), capsules (e.g.CS, ZC), pastes, pastilles, wettable powders or dusts (e.g. WP, SP, WS,DP, DS), pressings (e.g. BR, TB, DT), granules (e.g. WG, SG, GR, FG, GG,MG), insecticidal articles (e.g. LN), as well as gel formulations forthe treatment of plant propagation materials such as seeds (e.g. GF).These and further compositions types are defined in the “Catalogue ofpesticide formulation types and international coding system”, TechnicalMonograph No. 2, 6^(th) Ed. May 2008, CropLife International.

The mixtures of the invention can be formulated as an inoculant for aplant. The term “inoculant” means a composition that includes anisolated strain of the invention and optionally a carrier, which mayinclude a biologically acceptable medium.

Such inoculants and other suitable compositions can be prepared ascompositions comprising besides the active ingredients at least oneauxiliary (inert ingredient) by usual means (see e. g. H. D. Burges:Formulation of Microbial Biopesticides, Springer, 1998).

To produce a dry formulation, bacterial cells, preferably spores can besuspended in a suitable dry carrier (e. g. clay). To produce a liquidformulation, cells, preferably spores, can be re-suspended in a suitableliquid carrier (e. g. water-based) to the desired spore density. Thespore density number of spores per ml can be determined by identifyingthe number of colony-forming units (CFU) on agar medium e. g. potatodextrose agar after incubation for several days at temperatures of about20 to about 30° C.

According to one embodiment, individual components of the compositionaccording to the invention such as parts of a kit or parts of a binaryor ternary mixture may be mixed by the user himself in a spray tank orany other kind of vessel used for applications (e.g seed treater drums,seed pelleting machinery, knapsack sprayer) and further auxiliaries maybe added, if appropriate. When living microorganisms, such as thePaenibacillus strains of the invention, form part of such kit, it mustbe taken care that choice and amounts of the other parts of the kit (e.g. chemical pesticidal agents) and of the further auxiliaries should notinfluence the viability of the microbial pesticides in the compositionmixed by the user. Especially for bactericides and solvents,compatibility with the respective microbial pesticide has to be takeninto account.

The Paenibacillus strains as defined herein, whole culture broths,cell-free extracts, culture media and/or fusaricidins of formula I,together with the at least one biopesticide can be converted intocustomary types of agrochemical compositions, e. g. solutions,emulsions, suspensions, dusts, powders, pastes, granules, pressings,capsules, and mixtures thereof. Examples for composition types aresuspensions (e. g. SC, OD, FS), emulsifiable concentrates (e. g. EC),emulsions (e. g. EW, EO, ES, ME), capsules (e. g. CS, ZC), pastes,pastilles, wettable powders or dusts (e. g. WP, SP, WS, DP, DS),pressings (e. g. BR, TB, DT), granules (e. g. WG, SG, GR, FG, GG, MG),insecticidal articles (e. g. LN), as well as gel formulations for thetreatment of plant propagation materials such as seeds (e. g. GF). Theseand further compositions types are defined in the “Catalogue ofpesticide formulation types and international coding system”, TechnicalMonograph No. 2, 6^(th) Ed. May 2008, CropLife International.

The compositions are prepared in a known manner, such as described byMollet and Grubennann, Formulation technology, Wiley VCH, Weinheim,2001; or Knowles, New developments in crop protection productformulation, Agrow Reports DS243, T&F Informa, London, 2005.

Suitable auxiliaries are solvents, liquid carriers, solid carriers orfillers, surfactants, dispersants, emulsifiers, wetters, adjuvants,solubilizers, penetration enhancers, protective colloids, adhesionagents, thickeners, humectants, repellents, attractants, feedingstimulants, compatibilizers, bactericides, anti-freezing agents,anti-foaming agents, colorants, tackifiers and binders.

Suitable solvents and liquid carriers are water and organic solvents,such as mineral oil fractions of medium to high boiling point, e. g.kerosene, diesel oil; oils of vegetable or animal origin; aliphatic,cyclic and aromatic hydrocarbons, e. g. toluene, paraffin,tetrahydronaphthalene, alkylated naphthalenes; alcohols, e. g. ethanol,propanol, butanol, benzylalcohol, cyclohexanol; glycols; DMSO; ketones,e. g. cyclohexanone; esters, e. g. lactates, carbonates, fatty acidesters, gamma-butyrolactone; fatty acids; phosphonates; amines; amides,e. g. N-methylpyrrolidone, fatty acid dimethylamides; and mixturesthereof.

Suitable solid carriers or fillers are mineral earths, e. g. silicates,silica gels, talc, kaolins, limestone, lime, chalk, clays, dolomite,diatomaceous earth, bentonite, calcium sulfate, magnesium sulfate,magnesium oxide; polysaccharides, e. g. cellulose, starch; fertilizers,e. g. ammonium sulfate, ammonium phosphate, ammonium nitrate, ureas;products of vegetable origin, e. g. cereal meal, tree bark meal, woodmeal, nutshell meal, and mixtures thereof.

Suitable surfactants are surface-active compounds, such as anionic,cationic, nonionic and amphoteric surfactants, block polymers,polyelectrolytes, and mixtures thereof. Such surfactants can be used asemusifier, dispersant, solubilizer, wetter, penetration enhancer,protective colloid, or adjuvant. Examples of surfactants are listed inMcCutcheon's, Vol. 1: Emulsifiers & Detergents, McCutcheon'sDirectories, Glen Rock, USA, 2008 (International Ed. or North AmericanEd.).

Suitable anionic surfactants are alkali, alkaline earth or ammoniumsalts of sulfonates, sulfates, phosphates, carboxylates, and mixturesthereof. Examples of sulfonates are alkylarylsulfonates,diphenylsulfonates, alpha-olefin sulfonates, lignine sulfonates,sulfonates of fatty acids and oils, sulfonates of ethoxylatedalkylphenols, sulfonates of alkoxylated arylphenols, sulfonates ofcondensed naphthalenes, sulfonates of dodecyl- and tridecylbenzenes,sulfonates of naphthalenes and alkylnaphthalenes, sulfosuccinates orsulfosuccinamates. Examples of sulfates are sulfates of fatty acids andoils, of ethoxylated alkylphenols, of alcohols, of ethoxylated alcohols,or of fatty acid esters. Examples of phosphates are phosphate esters.Examples of carboxylates are alkyl carboxylates, and carboxylatedalcohol or alkylphenol ethoxylates.

Suitable nonionic surfactants are alkoxylates, N-subsituted fatty acidamides, amine oxides, esters, sugar-based surfactants, polymericsurfactants, and mixtures thereof. Examples of alkoxylates are compoundssuch as alcohols, alkylphenols, amines, amides, arylphenols, fatty acidsor fatty acid esters which have been alkoxylated with 1 to 50equivalents. Ethylene oxide and/or propylene oxide may be employed forthe alkoxylation, preferably ethylene oxide. Examples of N-subsitituedfatty acid amides are fatty acid glucamides or fatty acid alkanolamides.Examples of esters are fatty acid esters, glycerol esters ormonoglycerides. Examples of sugar-based surfactants are sorbitans,ethoxylated sorbitans, sucrose and glucose esters oralkylpolyglucosides. Examples of polymeric surfactants are home- orcopolymers of vinylpyrrolidone, vinyl alcohols, or vinyl acetate.

Suitable cationic surfactants are quaternary surfactants, for examplequaternary ammonium compounds with one or two hydrophobic groups, orsalts of long-chain primary amines. Suitable amphoteric surfactants arealkylbetains and imidazolines. Suitable block polymers are blockpolymers of the A-B or A-B-A type comprising blocks of polyethyleneoxide and polypropylene oxide, or of the A-B-C type comprising alkanol,polyethylene oxide and polypropylene oxide. Suitable polyelectrolytesare polyacids or polybases. Examples of polyacids are alkali salts ofpolyacrylic acid or polyacid comb polymers. Examples of polybases arepolyvinyl amines or polyethyleneannines.

Suitable adjuvants are compounds, which have a negligible or even nopesticidal activity themselves, and which improve the biologicalperformance of cell-free extract, culture medium or metabolite on thetarget. Examples are surfactants, mineral or vegetable oils, and otherauxilaries. Further examples are listed by Knowles, Adjuvants andadditives, Agrow Reports DS256, T&F Informa UK, 2006, chapter 5.

Suitable thickeners are polysaccharides (e. g. xanthan gum,carboxymethyl cellulose), inorganic clays (organically modified orunmodified), polycarboxylates, and silicates.

Suitable bactericides are bronopol and isothiazolinone derivatives suchas alkylisothiazolinones and benzisothiazolinones. Suitableanti-freezing agents are ethylene glycol, propylene glycol, urea andglycerin. Suitable anti-foaming agents are silicones, long chainalcohols, and salts of fatty acids. Suitable colorants (e. g. in red,blue, or green) are pigments of low water solubility and water-solubledyes. Examples are inorganic colorants (e. g. iron oxide, titan oxide,iron hexacyanoferrate) and organic colorants (e. g. alizarin-, azo- andphthalocyanine colorants). Suitable tackifiers or binders are polyvinylpyrrolidones, polyvinyl acetates, polyvinyl alcohols, polyacrylates,biological or synthetic waxes, and cellulose ethers.

When living microorganisms, such as bacterial strains of the genusPaenibacillus in form of cells or spores, form part of the compositions,such compositions can be prepared as compositions comprising besides theactive ingredients at least one auxiliary (inert ingredient) by usualmeans (see e. g. H. D. Burges: Formulation of Microbial Biopesticides,Springer, 1998). Suitable customary types of such compositions aresuspensions, dusts, powders, pastes, granules, pressings, capsules, andmixtures thereof. Examples for composition types are suspensions (e. g.SC, OD, FS), capsules (e. g. CS, ZC), pastes, pastilles, wettablepowders or dusts (e. g. WP, SP, WS, DP, DS), pressings (e. g. BR, TB,DT), granules (e. g. WG, SG, GR, FG, GG, MG), insecticidal articles (e.g. LN), as well as gel formulations for the treatment of plantpropagation materials such as seeds (e. g. GF). Herein, it has to betaken into account that each formulation type or choice of auxiliaryshould not influence the viability of the microorganism during storageof the composition and when finally applied to the soil, plant or plantpropagation material. Suitable formulations are e. g. mentioned in WO2008/002371, U.S. Pat. Nos. 6,955,912, 5,422,107.

Examples for suitable auxiliaries are those mentioned earlier herein,wherein it must be taken care that choice and amounts of suchauxiliaries should not influence the viability of the microbialpesticides in the composition. Especially for bactericides and solvents,compatibility with the respective microorganism of the respectivemicrobial pesticide has to be taken into account. In addition,compositions with microbial pesticides may further contain stabilizersor nutrients and UV protectants. Suitable stabilizers or nutrients aree. g. alpha-tocopherol, trehalose, glutamate, potassium sorbate, varioussugars like glucose, sucrose, lactose and maltodextrine (H. D. Burges:Formulation of Microbial Biopesticides, Springer, 1998). Suitable UVprotectants are e. g. inorganic compounds like titanium dioxide, zincoxide and iron oxide pigments or organic compounds like benzophenones,benzotriazoles and phenyltriazines. The compositions may in addition toauxiliaries mentioned for compositions herein optionally comprise0.1-80% stabilizers or nutrients and 0.1-10% UV protectants.

The agrochemical compositions generally comprise between 0.01 and 95%,preferably between 0.1 and 90%, and in particular between 0.5 and 75%,by weight of active substance. The active substances are employed in apurity of from 90% to 100%, preferably from 95% to 100% (according toNMR spectrum).

Examples for composition types and their preparation are:

i) Water-Soluble Concentrates (SL, LS)

10-60 wt % of a mixture of the invention and 5-15 wt % wetting agent (e.g. alcohol alkoxylates) are dissolved in water and/or in a water-solublesolvent (e. g. alcohols) ad 100 wt %. The active substance dissolvesupon dilution with water.

ii) Dispersible Concentrates (DC)

5-25 wt % of a mixture of the invention and 1-10 wt % dispersant (e. g.polyvinyl pyrrolidone) are dissolved in organic solvent (e. g.cyclohexanone) ad 100 wt %. Dilution with water gives a dispersion.

iii) Emulsifiable Concentrates (EC)

15-70 wt % of a mixture of the invention and 5-10 wt % emulsifiers (e.g. calcium dodecylbenzenesulfonate and castor oil ethoxylate) aredissolved in water-insoluble organic solvent (e. g. aromatichydrocarbon) ad 100 wt %. Dilution with water gives an emulsion.

iv) Emulsions (EW, EO, ES)

5-40 wt % of a mixture of the invention and 1-10 wt % emulsifiers (e. g.calcium dodecylbenzenesulfonate and castor oil ethoxylate) are dissolvedin 20-40 wt % water-insoluble organic solvent (e. g. aromatichydrocarbon). This mixture is introduced into water ad 100 wt % by meansof an emulsifying machine and made into a homogeneous emulsion. Dilutionwith water gives an emulsion.

v) Suspensions (SC, OD, FS)

In an agitated ball mill, 20-60 wt % of a mixture of the invention arecomminuted with addition of 2-10 wt % dispersants and wetting agents (e.g. sodium lignosulfonate and alcohol ethoxylate), 0.1-2 wt % thickener(e. g. xanthan gum) and water ad 100 wt % to give a fine activesubstance suspension. Dilution with water gives a stable suspension ofthe active substance. For FS type composition up to 40 wt % binder (e.g. polyvinyl alcohol) is added.

vi) Water-Dispersible Granules and Water-Soluble Granules (WG, SG)

50-80 wt % of a mixture of the invention are ground finely with additionof dispersants and wetting agents (e. g. sodium lignosulfonate andalcohol ethoxylate) ad 100 wt % and prepared as water-dispersible orwater-soluble granules by means of technical appliances (e. g.extrusion, spray tower, fluidized bed). Dilution with water gives astable dispersion or solution of the active substance.

vii) Water-Dispersible Powders and Water-Soluble Powders (WP, SP, WS)

50-80 wt % of a mixture of the invention are ground in a rotor-statormill with addition of 1-5 wt % dispersants (e. g. sodiumlignosulfonate), 1-3 wt % wetting agents (e. g. alcohol ethoxylate) andsolid carrier (e. g. silica gel) ad 100 wt %. Dilution with water givesa stable dispersion or solution of the active substance.

viii) Gel (GW, GF)

In an agitated ball mill, 5-25 wt % of a mixture of the invention arecomminuted with addition of 3-10 wt % dispersants (e. g. sodiumlignosulfonate), 1-5 wt % thickener (e. g. carboxymethyl cellulose) andwater ad 100 wt % to give a fine suspension of the active substance.Dilution with water gives a stable suspension of the active substance.

ix) Microemulsion (ME)

5-20 wt % of a mixture of the invention are added to 5-30 wt % organicsolvent blend (e. g. fatty acid dimethylamide and cyclohexanone), 10-25wt % surfactant blend (e. g. alcohol ethoxylate and arylphenolethoxylate), and water ad 100%. This mixture is stirred for 1 h toproduce spontaneously a thermodynamically stable microemulsion.

x) Microcapsules (CS)

An oil phase comprising 5-50 wt % of a mixture of the invention, 0-40 wt% water insoluble organic solvent (e. g. aromatic hydrocarbon), 2-15 wt% acrylic monomers (e. g. methylmethacrylate, methacrylic acid and a di-or triacrylate) are dispersed into an aqueous solution of a protectivecolloid (e. g. polyvinyl alcohol). Radical polymerization initiated by aradical initiator results in the formation of poly(meth)acrylatemicrocapsules.

Alternatively, an oil phase comprising 5-50 wt % of a whole culturebroth, cell-free extract, culture medium or metabolite of the invention,0-40 wt % water insoluble organic solvent (e. g. aromatic hydrocarbon),and an isocyanate monomer (e. g. diphenylmethene-4,4′-diisocyanatae) aredispersed into an aqueous solution of a protective colloid (e. g.polyvinyl alcohol). The addition of a polyamine (e. g.hexamethylenediamine) results in the formation of polyureamicrocapsules. The monomers amount to 1-10 wt %. The wt % relate to thetotal CS composition.

xi) Dustable Powders (DP, DS)

1-10 wt % of a whole culture broth, cell-free extract, culture medium ormetabolite of the invention are ground finely and mixed intimately withsolid carrier (e. g. finely divided kaolin) ad 100 wt %.

xii) Granules (GR, FG)

0.5-30 wt % of a whole culture broth, cell-free extract, culture mediumor metabolite of the invention are ground finely and associated withsolid carrier (e. g. silicate) ad 100 wt %. Granulation is achieved byextrusion, spray-drying or fluidized bed.

xiii) Ultra-Low Volume Liquids (UL)

1-50 wt % of a whole culture broth, cell-free extract, culture medium ormetabolite of the invention are dissolved in organic solvent (e. g.aromatic hydrocarbon) ad 100 wt %.

The compositions types i) to xiii) may optionally comprise furtherauxiliaries, such as 0.1-1 wt % bactericides, 5-15 wt % anti-freezingagents, 0.1-1 wt % anti-foaming agents, and 0.1-1 wt % colorants.

Solutions for seed treatment (LS), suspoemulsions (SE), flowableconcentrates (FS), powders for dry treatment (DS), water-dispersiblepowders for slurry treatment (WS), water-soluble powders (SS), emulsions(ES), emulsifiable concentrates (EC) and gels (GF) are usually employedfor the purposes of treatment of plant propagation materials,particularly seeds.

Preferred examples of seed treatment formulation types or soilapplication for pre-mix compositions are of WS, LS, ES, FS, WG orCS-type.

Typically, a pre-mix formulation for seed treatment applicationcomprises 0.5 to 99.9 percent, especially 1 to 95 percent, of thedesired ingredients, and 99.5 to 0.1 percent, especially 99 to 5percent, of a solid or liquid adjuvant (including, for example, asolvent such as water), where the auxiliaries can be a surfactant in anamount of 0 to 50 percent, especially 0.5 to 40 percent, based on thepre-mix formulation. Whereas commercial products will preferably beformulated as concentrates (e. g., pre-mix composition (formulation)),the end user will normally employ dilute formulations (e. g., tank mixcomposition).

Seed treatment methods for applying or treating the strains, wholeculture broths, cell-free extracts, culture media, fusaricidins offormula I and compositions of the invention, respectively, to plantpropagation material, especially seeds, are known in the art, andinclude dressing, coating, filmcoating, pelleting and soakingapplication methods of the propagation material. Such methods are alsoapplicable to the combinations according to the invention. In apreferred embodiment, the mixtures and compositions of the invention,respectively, are applied or treated onto the plant propagation materialby a method such that the germination is not negatively impacted.Accordingly, examples of suitable methods for applying (or treating) aplant propagation material, such as a seed, is seed dressing, seedcoating or seed pelleting and alike.

It is preferred that the plant propagation material is a seed, seedpiece (i.e. stalk) or seed bulb.

Although it is believed that the present method can be applied to a seedin any physiological state, it is preferred that the seed be in asufficiently durable state that it incurs no damage during the treatmentprocess. Typically, the seed would be a seed that had been harvestedfrom the field; removed from the plant; and separated from any cob,stalk, outer husk, and surrounding pulp or other non-seed plantmaterial. The seed would preferably also be biologically stable to theextent that the treatment would cause no biological damage to the seed.It is believed that the treatment can be applied to the seed at any timebetween harvest of the seed and sowing of the seed or during the sowingprocess (seed directed applications). The seed may also be primed eitherbefore or after the treatment.

Even distribution of the ingredients in the mixtures and compositions ofthe invention, respectively, and adherence thereof to the seeds isdesired during propagation material treatment. Treatment could vary froma thin film (dressing) of the formulation containing the combination,for example, a mixture of active ingredient(s), on a plant propagationmaterial, such as a seed, where the original size and/or shape arerecognizable to an intermediary state (such as a coating) and then to athicker film (such as pelleting with many layers of different materials(such as carriers, for example, clays; different formulations, such asof other active ingredients; polymers; and colourants) where theoriginal shape and/or size of the seed is no longer recognizable.

An aspect of the present invention includes application of the mixturesand compositions of the invention, respectively, onto the plantpropagation material in a targeted fashion, including positioning theingredients in the combination onto the entire plant propagationmaterial or on only parts thereof, including on only a single side or aportion of a single side. One of ordinary skill in the art wouldunderstand these application methods from the description provided inEP954213B1 and WO06/112700.

The strains, whole culture broths, cell-free extracts, culture media,fusaricidins of formula I and compositions of the invention,respectively, can also be used in form of a “pill” or “pellet” or asuitable substrate and placing, or sowing, the treated pill, orsubstrate, next to a plant propagation material. Such techniques areknown in the art, particularly in EP1124414, WO07/67042, and WO07/67044. Application of the strains, whole culture broths, cell-freeextracts, culture media, fusaricidins of formula I and compositions,respectively, described herein onto plant propagation material alsoincludes protecting the plant propagation material treated with thecombination of the present invention by placing one or morepesticide-containing particles next to a pesticide-treated seed, whereinthe amount of pesticide is such that the pesticide-treated seed and thepesticide-containing particles together contain an effective dose of thepesticide and the pesticide dose contained in the pesticide-treated seedis less than or equal to the maximal non-phytotoxic dose of thepesticide. Such techniques are known in the art, particularly inWO2005/120226.

Application of the strains, whole culture broths, cell-free extracts,culture media, fusaricidins of formula I and compositions of theinvention, respectively, onto the seed also includes controlled releasecoatings on the seeds, wherein the ingredients of the combinations areincorporated into materials that release the ingredients over time.Examples of controlled release seed treatment technologies are generallyknown in the art and include polymer films, waxes, or other seedcoatings, wherein the ingredients may be incorporated into thecontrolled release material or applied between layers of materials, orboth.

Seed can be treated by applying thereto the mixtures and compositions ofthe invention, respectively, in any desired sequence or simultaneously.

The seed treatment occurs to an unsown seed, and the term “unsown seed”is meant to include seed at any period between the harvest of the seedand the sowing of the seed in the ground for the purpose of germinationand growth of the plant.

Treatment to an unsown seed is not meant to include those practices inwhich the active ingredient is applied to the soil but would include anyapplication practice that would target the seed during the plantingprocess.

Preferably, the treatment occurs before sowing of the seed so that thesown seed has been pre-treated with the mixtures and compositions of theinvention, respectively. In particular, seed coating or seed pelletingare preferred. As a result of the treatment, the ingredients are adheredon to the seed and therefore available for pest control.

The treated seeds can be stored, handled, sowed and tilled in the samemanner as any other active ingredient treated seed.

In particular, the present invention relates to a method for protectionof plant propagation material from pests, harmful fungi and/or improvingthe health of plants grown from said plant propagation material, whereinthe soil, wherein plant propagation material is sown, is treated with aneffective amount of a mixture or composition of the invention,respectively.

The user applies the compositions of the invention usually from apredosage device, a knapsack sprayer, a spray tank, a spray plane, or anirrigation system. Usually, the agrochemical composition is made up withwater, buffer, and/or further auxiliaries to the desired applicationconcentration and the ready-to-use spray liquor or the agrochemicalcomposition according to the invention is thus obtained. Usually, 20 to2000 liters, preferably 50 to 400 liters, of the ready-to-use sprayliquor are applied per hectare of agricultural useful area.

When it comes to the treatment of plant propagation material, especiallyseeds, the compositions disclosed herein give, after two-to-tenfolddilution, active components concentrations of from 0.01 to 60% byweight, preferably from 0.1 to 40%, in the ready-to-use preparations.Application can be carried out before or during sowing. Methods forapplying a strain, cell-free extract, culture medium, metabolite orcomposition of the invention, respectively, onto plant propagationmaterial, especially seeds, include dressing, coating, pelleting,dusting, soaking and in-furrow application methods of the propagationmaterial. Preferably, the strains, whole culture broths, cell-freeextracts, culture media, fusaricidins of formula I or compositions ofthe invention, respectively, are applied onto the plant propagationmaterial by a method such that germination is not induced, e. g. by seeddressing, pelleting, coating and dusting.

When the bacterial strains are employed in crop protection, wherein thestrains are applied as foliar treatment or to the soil, the applicationrates usually range from about 1×10⁶ to 5×10¹⁶ (or more) CFU/ha,preferably from about 1×10⁷ to about 1×10¹⁶ CFU/ha, even more preferablyfrom 1×10⁹ to 5×10¹⁵ CFU/ha and particularly preferred even morepreferably from 1×10¹² to 5×10¹⁴ CFU/ha. In the case of(entomopathogenic) nematodes as microbial pesticides (e. g. Steinernemafeltiae), the application rates preferably range inform about 1×10⁵ to1×10¹² (or more), more preferably from 1×10⁸ to 1×10¹¹, even morepreferably from 5×10⁸ to 1×10¹⁰ individuals (e. g. in the form of eggs,juvenile or any other live stages, preferably in an infetive juvenilestage) per ha.

When the strains of the invention are employed in seed treatment, theapplication rates with respect to plant propagation material usuallyrange from about 1×10¹ to 1×10¹² (or more) CFU/seed, preferably fromabout 1×10³ to about 1×10¹⁰ CFU/seed, and even more preferably fromabout 1×10³ to about 1×10⁶ CFU/seed. Alternatively, the applicationrates with respect to plant propagation material preferably range fromabout 1×10⁷ to 1×10¹⁶ (or more) CFU per 100 kg of seed, preferably from1×10⁹ to about 1×10¹⁵ CFU per 100 kg of seed, even more preferably from1×10¹¹ to about 1×10¹⁵ CFU per 100 kg of seed.

When cell-free extracts, culture media and/or metabolites such asfusaricidins of formula I are employed, the solid material (dry matter)are considered as active components, e. g. to be obtained after dryingor evaporation of the extraction medium or the suspension medium in caseof liquid formulations. When employed in plant protection, the amountsof active components applied are, depending on the kind of effectdesired, from 0.001 to 2 kg per ha, preferably from 0.005 to 2 kg perha, more preferably from 0.05 to 0.9 kg per ha, and in particular from0.1 to 0.75 kg per ha. In treatment of plant propagation materials suchas seeds, e. g. by dusting, coating or drenching seed, amounts of activecomponents of from 0.1 to 1000 g, preferably from 1 to 1000 g, morepreferably from 1 to 100 g and most preferably from 5 to 100 g, per 100kilogram of plant propagation material (preferably seeds) are generallyrequired. When used in the protection of materials or stored products,the amount of active components applied depends on the kind ofapplication area and on the desired effect. Amounts customarily appliedin the protection of materials are 0.001 g to 2 kg, preferably 0.005 gto 1 kg, of active components per cubic meter of treated material.

According to one embodiment, individual components of the composition ofthe invention such as parts of a kit or parts of a binary or ternarymixture may be mixed by the user himself in a spray tank or any otherkind of vessel used for applications (e.g seed treater drums, seedpelleting machinery, knapsack sprayer) and further auxiliaries may beadded, if appropriate.

If living microorganisms, such as the bacterial strains of the genusPaenibacillus, form part of such kit, it must be taken care that choiceand amounts of the components (e. g. chemical pesticidal agents) and ofthe further auxiliaries should not influence the viability of themicroorganisms in the composition mixed by the user. Especially forbactericides and solvents, compatibility with the respectivemicroorganisms has to be taken into account.

Consequently, one embodiment of the invention is a kit for preparing ausable pesticidal composition, the kit comprising a) a compositioncomprising component 1) as defined herein and at least one auxiliary;and b) a composition comprising component 2) as defined herein and atleast one auxiliary; and optionally c) a composition comprising at leastone auxiliary and optionally a further active component 3) as definedherein.

Various types of oils, wetters, adjuvants, fertilizer, ormicronutrients, and further pesticides (e. g. herbicides, insecticides,fungicides, growth regulators, safeners, biopesticides) may be added tothe mixtures and compositions of the invention, respectively as premixor, if appropriate not until immediately prior to use (tank mix). Theseagents can be admixed with the compositions according to the inventionin a weight ratio of 1:100 to 100:1, preferably 1:10 to 10:1.Preferably, a composition of the invention comprises aa third activecomponent a further biopesticide. Even more preferably, a composition ofthe invention comprises besides an auxiliary and at least onefusaricidins of formula I and a biopesticide II as defined herein, amicrobial pesticide.

A pesticide is generally a chemical or biological agent (such as avirus, bacterium, antimicrobial or disinfectant) that through its effectdeters, incapacitates, kills or otherwise discourages pests. Targetpests can include insects, plant pathogens, weeds, mollusks, birds,mammals, fish, nematodes (roundworms), and microbes that destroyproperty, cause nuisance, spread disease or are vectors for disease. Theterm pesticides includes also plant growth regulators that alter theexpected growth, flowering, or reproduction rate of plants; defoliantsthat cause leaves or other foliage to drop from a plant, usually tofacilitate harvest; desiccants that promote drying of living tissues,such as unwanted plant tops; plant activators that activate plantphysiology for defense of against certain pests; safeners that reduceunwanted herbicidal action of pesticides on crop plants; and plantgrowth promoters that affect plant physiology to increase plant growth,biomass, yield or any other quality parameter of the harvestable goodsof a crop plant.

By applying at least one Paenibacillus strain as defined in any one ofthe preferred embodiments above, or the culture medium or a cell-freeextract thereof or at least one metabolite thereof together with atleast one biopesticide II from groups L1) to L5) a synergistic effectcan be obtained, i.e. more then simple addition of the individualeffects is obtained (synergistic mixtures).

According to one embodiment, the mixtures comprise component 1) andcomponent 2) in a synergistically effective amount.

The term “synergstic effect” is understood to refer in particular tothat defined by Colby's formula (Colby, S. R., “Calculating synergisticand antagonistic responses of herbicide combinations”, Weeds, 15, pp.20-22, 1967).

The term “synergistic effect” is also understood to refer to thatdefined by application of the Tammes method, (Tammes, P. M. L.,“Isoboles, a graphic representation of synergism in pesticides”,Netherl. J. Plant Pathol. 70, 1964).

This can be obtained by applying at least one Paenibacillus strain asdefined in any one of the preferred embodiments above, or the culturemedium or a cell-free extract thereof or at least one metabolite thereofand at least one pesticide II simultaneously, either jointly (e. g. astank-mix) or seperately, or in succession, wherein the time intervalbetween the individual applications is selected to ensure that theactive substance applied first still occurs at the site of action in asufficient amount at the time of application of the further activesubstance(s). The order of application is not essential for working ofthe present invention.

When applying a Paenibacillus strain as defined in any one of thepreferred embodiments above, or the culture medium or a cell-freeextract thereof or a metabolite thereof and a pesticide II sequentiallythe time between both applications may vary e. g. between 2 hours to 7days. Also a broader range is possible ranging from 0.25 hour to 30days, preferably from 0.5 hour to 14 days, particularly from 1 hour to 7days or from 1.5 hours to 5 days, even more preferred from 2 hours to 1day. It is preferred that the microbial pesticides (e.g. Paenibacillusstrains of component 1) and/or microbial pesticides from groups L1), L3)and L5)) are applied as last treatment.

According to the invention, the solid material (dry matter) of thebiopesticides (with the exception of oils such as Neem oil) areconsidered as active components (e. g. to be obtained after drying orevaporation of the extraction or suspension medium in case of liquidformulations of the microbial pesticides).

In accordance with the present invention, the weight ratios andpercentages used herein for a biological extract such as Quillay extractare based on the total weight of the dry content (solid material) of therespective extract(s).

The total weight ratios of compositions comprising at least onemicrobial pesticide in the form of viable microbial cells includingdormant forms, can be determined using the amount of CFU of therespective microorganism to calculate the total weight of the respectiveactive component with the following equation that 1×10¹⁰ CFU equals onegram of total weight of the respective active component. Colony formingunit is measure of viable microbial cells, in particular fungal andbacterial cells. In addition, here “CFU” may also be understood as thenumber of (juvenile) individual nematodes in case of (entomopathogenic)nematode biopesticides, such as Steinernema feltiae.

In the mixtures and compositions according to the invention the weightratio of the component 1) and the component 2) generally depends fromthe properties of the active components used, usually it is in the rangeof from 1:10,000 to 10,000:1, often it is in the range of from 1:100 to100:1, regularly in the range of from 1:50 to 50:1, preferably in therange of from 1:20 to 20:1, more preferably in the range of from 1:10 to10:1, even more preferably in the range of from 1:4 to 4:1 and inparticular in the range of from 1:2 to 2:1.

According to further embodiments of the mixtures and compositions, theweight ratio of the component 1) and the component 2) usually is in therange of from 1000:1 to 1:1, often in the range of from 100:1 to 1:1,regularly in the range of from 50:1 to 1:1, preferably in the range offrom 20:1 to 1:1, more preferably in the range of from 10:1 to 1:1, evenmore preferably in the range of from 4:1 to 1:1 and in particular in therange of from 2:1 to 1:1.

According to further embodiments of the mixtures and compositions, theweight ratio of the component 1) and the component 2) usually is in therange of from 20,000:1 to 1:10, often in the range of from 10,000:1 to1:1, regularly in the range of from 5,000:1 to 5:1, preferably in therange of from 5,000:1 to 10:1, more preferably in the range of from2,000:1 to 30:1, even more preferably in the range of from 2,000:1 to100:1 and in particular in the range of from 1,000:1 to 100:1.

According to further embodiments of the mixtures and compositions, theweight ratio of the component 1) and the component 2) usually is in therange of from 1:1 to 1:1000, often in the range of from 1:1 to 1:100,regularly in the range of from 1:1 to 1:50, preferably in the range offrom 1:1 to 1:20, more preferably in the range of from 1:1 to 1:10, evenmore preferably in the range of from 1:1 to 1:4 and in particular in therange of from 1:1 to 1:2.

According to further embodiments of the mixtures and compositions, theweight ratio of the component 1) and the component 2) usually is in therange of from 10:1 to 1:20,000, often in the range of from 1:1 to1:10,000, regularly in the range of from 1:5 to 1:5,000, preferably inthe range of from 1:10 to 1:5,000, more preferably in the range of from1:30 to 1:2,000, even more preferably in the range of from 1:100 to1:2,000 to and in particular in the range of from 1:100 to 1:1,000.

The mixtures and compositions thereof according to the invention can, inthe use form as fungicides and/or insecticides, also be present togetherwith other active substances, e. g. with herbicides, insecticides,growth regulators, fungicides or else with fertilizers, as pre-mix or,if appropriate, not until immeadiately prior to use (tank mix).

Mixing the binary mixtures of the invention or the compositionscomprising them with other fungicides i results in many cases in anexpansion of the fungicidal spectrum of activity or in a prevention offungicide resistance development. Furthermore, in many cases,synergistic effects are obtained.

Mixing the binary mixtures of the invention or the compositionscomprising them with other insecticides results in many cases in anexpansion of the insecticidal spectrum of activity or in a prevention ofinsecticide resistance development. Furthermore, in many cases,synergistic effects are obtained.

According to the present invention, it may be preferred that themixtures and compsiitons comprising them, comprise besides at least onePaenibacillus strain as defined in any one of the preferred embodimentsabove, the culture medium or a cell-free extract thereof or at least onemetabolite thereof (component 1), and a biopesticide II (component 2),as component 3) a further pesticide, preferably in a synergisticallyeffective amount. Another embodiment relates to mixtures wherein thecomponent 3) is a pesticide III selected from groups SF) and SI) asdefined below. These ternary mixtures are especially suitable fortreatment of plant propagation materials (i.e. seed treatment).

The following list of pesticides III, in conjunction with which thebinary mixtures according to the invention can be used, is intended toillustrate the possible combinations but does not limit them:

SF) Fungicides

-   -   inhibitors of complex III at Q_(o) site selected from:        pyraclostrobin, azoxystrobin, picoxystrobin, trifloxystrobin,        dimoxystrobin, enestroburin, fenaminstrobin, fluoxastrobin,        kresoxim-methyl, mandestrobine, metominostrobin, orysastrobin,        pyrametostrobin, pyraoxystrobin;    -   broad-spectrum pyridine and pyrazole inhibitors of complex II        selected from: fluxapyroxad, boscalid, benzovindiflupyr,        penflufen, penthiopyrad, sedaxane, fluopyram, bixafen,        isopyrazam;    -   Basidiomycetes-specific inhibitors of complex II selected from:        carboxin, benodanil, fenfuram, flutolanil, furametpyr, mepronil,        oxycarboxin, thifluzamide; ATP production inhibitor silthiofam;    -   fungicidal azole compounds selected from: ipconazole,        difenoconazole, prothioconazole, prochloraz, triticonazole,        flutriafol, cyproconazole, diniconazole, diniconazole-M,        fluquinconazole, flusilazole, hexaconazole, imazalil,        imibenconazole, metconazole, myclobutanil, simeconazole,        tebuconazole, triadimenol, uniconazole, thiabendazole;    -   Oomycetes fungicides selected from: oxathiapiprolin,        valifenalate, metalaxyl, metalaxyl-M, ethaboxam, dimethomorph,        zoxamide, flumorph, mandipropamid, pyrimorph, benthiavalicarb,        iprovalicarb;    -   MAP/histidine kinase inhibitor: fludioxonil;    -   benzimidazole compounds selected from: thiophanate-methyl,        carbendazim;    -   dithiocarbamate compounds selected from: thiram, ziram;        SI) Insecticides    -   GABA antagonist compounds selected from: fipronil, ethiprole,        vaniliprole, pyrafluprole, pyriprole,        5-amino-1-(2,6-dichloro-4-methyl-phenyl)-4-sulfinannoyl-1H-pyrazole-3-carbothioic        acid amide;    -   lepidopteran-specific ryanodine receptor inhibitors selected        from: chlorantraniliprole and flubendiamide;    -   cross-spectrum ryanodine receptor inhibitor: cyantraniliprole;    -   pyrethroid sodium channel modulators selected from: tefluthrin,        bifenthrin, cypermethrin, alpha-cypermethrin, cyfluthrin,        beta-cyfluthrin, lambda-cyhalothrin, deltamethrin,        esfenvalerate, etofenprox, fenvalerate, flucythrinate,        permethrin;    -   systemically-active neonicotinoid compounds: clothianidin,        imidacloprid, thiamethoxam, dinotefuran, acetamiprid,        flupyradifurone, thiacloprid, triflumezopyrim, nitenpyram;    -   Acetylcholinesterase inhibitors, chloride channel activators and        sulfoximines: sulfoxaflor, acephate, chlorpyrifos, thiodicarb,        abamectin, spinosad;    -   other insecticide: tioxazafen.

More preferably, the pesticides III are selected from the followinggroups SF) and SI):

SF) Fungicides

-   -   azoxystrobin, trifloxystrobin, picoxystrobin, pyraclostrobin,        sedaxane, penthiopyrad, penflufen, fluopyram, fluxapyroxad,        boscalid, oxathiapiprolin, metalaxyl, metalaxyl-M, ethaboxam,        dimethomorph, valifenalate, cyproconazole, cifenoconazole,        prothioconazole, flutriafol, thiabendazole, ipconazole,        tebuconazole, triadimenol, prochloraz, fluquinconazole,        triticonazole, fludioxinil, carboxin, silthiofam, ziram, thiram,        carbendazim, thiophanate-methyl;        SI) Insecticides    -   fipronil, clothianidin, thiamethoxam, acetamiprid, dinotefuran,        imidadoprid, thiacloprid, sulfoxaflor, methiocarb, tefluthrin,        bifenthrin, cypermethrin, alpha-cypermethrin, spinosad,        chlorantraniliprole, cyantraniliprole, thiodicarb,        triflumezopyrim, acephate, chlorpyriphos, flupyradifurone,        abamectin, tioxazafen.

EXAMPLES

The present invention will be described in greater detail by means ofthe following examples. The following examples are for illustrativepurposes and are not intended to limit the scope of the invention.

Examples Concerning Paenibacillus Strains and Metabolites ThereofExample 1: Isolation of Paenibacillus Strains

Soil samples from a variety of European locations including Germany werecollected. By applying commonly known microbial isolation procedures tothese soils, the inventors obtained a variety of bacteria that werefurther subjected to conventional isolation techniques for providingpure isolates as described herein.

Standard microbial enrichment technique (C. A. Reddy, T. J. Beveridge,J. A. Breznak, G. A. Marzluf, T. M. Schmidt, and L. R. Snyder (eds.).Methods for General and Molecular Microbiology, Am. Soc. Microbiol.,Washington, D.C.) was followed to isolate each type of bacteria.

The following strains have been isolated and deposited under BudapestTreaty with the Deutsche Sammlung von Mikroorganismen and Zellkulturen(DSMZ) on Feb. 20, 2013:

a) Lu16774 as deposited with DSMZ having the deposit number DSM 26969

b) Lu17007 as deposited with DSMZ having the deposit number DSM 26970

c) Lu17015 as deposited with DSMZ having the deposit number DSM 26971.

Example 2—Characterization of Paenibacillus Strains Example 2.1:16S-rDNA Sequencing

The 16S rRNA gene sequences of the Paenibacillus strains were determinedby direct sequencing of PCR-amplified 16S rDNA at the DSMZ,Braunschweig, Germany.

Genomic DNA extraction was carried out using the MasterPure™ GramPositive DNA Purification Kit from Epicentre Biotechnologies accordingto the manufacturer's instructions. PCR-mediated amplification of the16S rDNA and purification of the PCR product was carried out asdescribed previously (Int. J. Syst. Bacteriol. 46, 1088-1092, 1996).Purified PCR products were sequenced using the Big Dye® Terminator v1.1Cycle Sequencing Kit (Applied Biosystems) as directed in themanufacturers protocol. Sequence reactions were electrophoresed usingthe 3500xL Genetic Analyzer from Applied Biosystems. Sequenceambiguities may be due to the existence of several cistrons encoding 16SrRNAs with different sequences within a single genome (J. Bacteriol.178(19), 5636-5643, 1996).

The resulting sequence data from the strains was put into the alignmenteditor AE2 (http://iubio.bio.indiana.edu/soft/molbio/unix/ae2.readnne),aligned manually according to the secondary structure of the resultingrRNA molecule and compared with representative 16S rRNA gene sequencesof organisms belonging to the Firmicutes (Nucl. Acids Res. 27, 171-173,1999). For comparison, 16S rRNA sequences were obtained from the EMBLand RDP data bases.

The 16S rDNA sequences of the strains of the invention are set forth inthe Sequence Listing as indicated in Table 2.

TABLE 2 Sequence listing references of the 16S rDNA of the Paenibacillusstrains. Strain SEQ ID NO Lu16774 1 Lu17007 2 Lu17015 3

The 16S rDNA gene identity values in % were calculated by pairwisecomparison of the sequences within the alignment of the sequencescompared.

Comparison performed of only two sequences based on pairwise sequencealignment are denoted herein as binary values. The other values arebased on a multiple sequence alignment of all sequences within thecomparison. Higher identity values from multi-sequence comparisonsresult from the problem that the sequence data of the compared sequenceswere of different length resulting in a shorter alignment.

The % identity from pair-wise comparisons of the complete rDNA sequencesamong the three strains Lu16774, Lu17007 and Lu17015 was between 99.5and 99.9% (Table 3, binary values).

TABLE 3 Identity in % of the complete 16S rRNA sequences of threePaenibacillus strains (binary values in brackets). Identity of thecomplete 16S rRNA sequence of Paenibacillus strains (%) Strains Lu16774Lu17015 Lu17007 Lu16774 — Lu17015 99.7 (99.5) — Lu17007 99.9 (99.8) 99.8(99.5) —

The comparison of the complete 16S rRNA sequence of the three strainsLu16774, Lu17007 and Lu17015 with related taxa (see FIG. 9) revealed ahigh percentage of identity to Paenibacillus peoriae (type-strain DSM8320) with 99.8%. The binary values for pairwise-sequence alignments ofP. peoriae with the strains Lu16774, Lu17007 and Lu17015 were asfollows: Lu16774: 99.5%, Lu17007: 99.5%; and Lu17015: 99.7% identity,respectively.

A final evaluation of species to which the Paenibacillus strainsLu16774, Lu17015 and Lu17007 belong was based on the 16S rRNA sequencedata not possible.

The sequencing of the complete rDNA resulted for Paenibacillus peoriaeNRRL BD-62 in 100.0% identity to P. peoriae (type strain DSM 8320)confirming the species designation P. peoriae for this strain BD-62 (seeFIG. 9).

The close relationship of all three Paenibacillus strains Lu16774,Lu17007 and Lu17015 to P. peoriae was confirmed by the comparison withthe 16S rRNA sequence of P. peoriae strain BD-62 which resulted inidentity values of 99.8% (see FIG. 9).

For construction of the phylogenetic dendrogram operations of the ARBpackage (Nucl. Acids Res. 35, 7188-7196, 2007) were used: based on theevolutionary distance values the phylogenetic tree was constructed bythe neighbor-joining method (Jukes, T. H. & Cantor C. R. (1969).Evolution of protein molecules. In Mammalian protein metabolism, pp.21-132. Edited by H. N. Munro. New York: Academic press) using thecorrection of Jukes and Cantor (Mol. Biol. Evol. 4, 406-425, 1987). Theroot of the tree was determined by including the 16S rRNA gene sequenceof Cohnella thermotolerans into the analysis. The scale bar below thedendrogram indicates 1 nucleotide substitutions per 100 nucleotides. Theresults are given in FIG. 10.

The phylogenetic dendrogram of these sequences (FIG. 10) shows that thethree strains Lu16774, Lu17007 and Lu17015 are most-closely related toeach other and that their closest relative known to each of them was thePaenibacillus peoriae strain NRRL BD-62.

Example 2.2: RiboPrint-Analysis

Standardized, automated ribotyping is performed using the QualiconRiboPrintersystem. The RiboPrinter system combines molecular processingsteps for ribotyping in a stand-alone, automated instrument. Theprocedure includes cell lysis, digestion of chromosomal DNA withrestriction enzyme EcoRI, separation of fragments by electrophoresis,transfer of DNA fragments to a nylon membrane, hybridization to a probegenerated from the rrnB operon from E. coli, chemiluminescent detectionof the probe to the fragments containing rrn operon sequences, imagedetection and computerized analysis of RiboPrint patterns (FoodTechnology 50(1), 77-81, 1996; Proc. Natl. Acad. Sci. USA 92, 5229-5233,1995; Int. Journ. Syst. Bact. 44(3), 454-460, 1994).

Ribotyping has been executed by the DSMZ, Germany with the Paenibacillusstrains Lu16774, Lu17007 and Lu17015 in comparison to the P. peoriaestrain BD-62 using the restriction enzyme EcoRI. The resulting patternshave been compared using the Software of the RiboPrinter system, theintegrated DuPont Identification Library as well as the BioNumericsSoftware (Applied Maths, Belgium).

Similarity of all three strains Lu16774, Lu17007 and Lu17015 to BD-62was between 0.24 and 0.5 (FIG. 11). The three strains group in twogroups, first comprising Lu17015, whereas the second group comprises thestrains Lu16774 and Lu17007. None of the strains has a similarity higherthan 0.84 to any strain within the DuPont Identification Library and wastherefore not identified automatically.

The strain BD-62 has been identified as Paenibacillus peoriae based onthe entry DUP-13142 of the DuPont identification library (entry based onPaenibacillus peoriae DSM 8320).

Example 2.3: Morphological and Physiological Characterization

The strains were characterized at the DSMZ in analogy to methodsdescribed in Gordon, R. E., Haynes, W. C. & Pang. C. H.-N. (1973): TheGenus Bacillus, Agriculture Handbook no. 427. Washington DC: USDepartment of Agriculture. The results are given in Table 4.

TABLE 4 Characterization Data of the Paenibacillus strains of theinvention and comparison to known Paenibacillus peoriae strain NRRLBD-62. Paenibacillus strains Identification Lu16774 Lu17007 Lu17015BD-62 Characteristics cell form rod- rod- rod- rod- shaped shaped shapedshaped width [μm] 0.9-1.0 0.9-1.0 0.9-1.0 0.9-1.0 length [μm]   3 -> 5.0  3-5.0   3-5.0 2.5-5.0 ellipsoid spores + + + + swollensporangium + + + + Catalase + + + + Oxidase − − − − anaerobicgrowth + + + + VP reaction + + + + pH in VP-Medium 5.2 5.7 4.8 5.2maximum temperature positive growth at ° C. 40 40 40 40 negative growthat ° C. 50 50 50 50 Growth in: Medium pH 5.7 + + + + NaCl 2% + + + +NaCl 5% − − − − NaCl 7% − − − − Acid formation from: D-Glucose + + + +L-Arabinose + + + + D-Xylose + + + + D-Mannitol + + + +D-Fructose + + + + Raffinose + + + + Trehalose + + + − Glycerol + + + +Gas from glucose + + + + Hydrolysis of starch + + + + gelatin + + + +casein + + + ? Tween 80 − − − − esculin + + + + Utilisation of citraten.g.* n.g. n.g. n.g. propionate n.g. n.g. n.g. n.g. N0₃ to N0₂ + + + +Indole reaction − − − − Lecithinase + + + − Phenylalanine desaminase − −− − Arginine dihydrolase − − − − Lysozyme + + + + *n.g. = no growth.

Analysis of the cellular fatty acids performed at the DSMZ resulted thatall strains showed at typical profile for Paenibacillus spp.

Using the available genetic, physiological and biochemical data, it isshown that the strains Lu16774, Lu17007 and Lu17015 belong to the genusPaenibacillus. As the strains Lu16774, Lu17007 and Lu17015 as well asBD-62 do produce gas from glucose, none of them belongs to Paenibacillusjamilae.

A phenotypic differentiation between Paenibacillus peoriae andPaenibacillus polymyxa is primarily possible using characteristics ofacid production from certain substrates (Int. J. Syst. Bacteriol. 43(2),388-390, 1993; In. J. Syst. Bacteriol. 46(6), 988-1003, 1996). None ofthe strains Lu16774, Lu17007 and Lu17015 did completely match with itscharacteristics outlined in Table 4 completely to any of these twospecies, but in sum of the available genetic, physiological andbiochemical data most likely point to the species Paenibacillus peoriaeand P. polymyxa or at least to another species very closely related toPaenibacillus peoriae and P. polymyxa.

Due to the multitude of Paenibacillus species described so far, it isimpossible to determine the correct taxonomic species of the threeisolates tested based on physiological and morphological criteria fromTable 4 (Rainer Borriss, Humboldt University Berlin, unpublishedresults).

Nevertheless, it was not possible to completely determine the specieswithin this genus. The most closely related species and strain was foundto be Paenibacillus peoriae BD-62 based on 16S-rDNA analysis (see e. g.FIG. 11).

Example 2.4: Phylogenetic Analysis Based on Genes Coding for DnaN, GyrB,RecF, RecN and RpoA

The nucleotide sequences of the genes coding for DnaN, GyrB, RecF, RecNand RpoA have been extracted from complete genome sequences or frompublic databases (Sequence listings as outlined in Table 28).

The identity tables (FIGS. 12 to 16) have been generated with an allagainst all approach where every sequence is aligned with every othersequence. The sequence alignment was performed with a program needle(EMBOSS package 6.6.0; Trends in Genetics 16 (6), 276-277). Standardparameters where used (gap creation 10.0; gap extension 0.5). IdentityScores are are calculated on the basis of the alignments without takingany gaps into account.

For the phylogenetic trees (FIGS. 17 to 21), multiple sequencealignments that have been performed with Clustal Omega (version 1.2.0;Molecular Systems Biology 7: 539, doi:10.1038/msb.2011.75). Thephylogenetic trees are calculated by maximum likelyhood method with thesoftware Dnaml (implemented in the Phylip 3.696 package; Felsenstein1981, http://evolution.genetics.washington.edu/phylip.html). Thedendrograms have been established using a F84 distance model whileapplying a transition-transversion ratio of two (2). The trees areplotted with the tool Dendroscope (http://dendroscope.org/).

TABLE 28 Sequence listing references of the dnaN, gyrB, recF, recN andrpoA DNA sequences of the Paenibacillus strains. Strain Gene SEQ ID NOLu16774 dnaN 4 Lu16774 gyrB 5 Lu16774 recF 6 Lu16774 recN 7 Lu16774 rpoA8 Lu17007 dnaN 9 Lu17007 gyrB 10 Lu17007 recF 11 Lu17007 recN 12 Lu17007rpoA 13 Lu17015 dnaN 14 Lu17015 gyrB 15 Lu17015 recF 16 Lu17015 recN 17Lu17015 rpoA 18

Example 2.5: Core Genome Comparisons and AAI Matrix

Genome comparisons have be performed using the software package EDGAR ofthe university Gießen (BMC Bioinformatics 10, 154, 2009;(https://edgar.connputational.bio.unigiessen.de/cgi-bin/edgar.cgi). Thedetermination of the core genome, the phylogenetic dendrograms on thebasis of the complete genome sequences and the AAI matrix values havebeen performed using the software package EDGAR. Results are shown inFIG. 22.

Example 3: Growth (Fermentability) of Strains for In-Vivo Tests

For green-house and field trials, the Paenibacillus strains were firstgrown on ISP2 plates (ready-to-use agar from BD [USA], catalog number277010). Afterwards, baffled shake flasks containing liquid ISP2 mediumwere inoculated with a colony from the agar plate and incubated for 5-7days at 150 rpm and 25° C. Depending on the test, either whole culturebroth, or the centrifuged and H₂O-washed cell pellet, or the supernatantwas applied to the plants. A scale-up to 10 L fermenters was possible.

Paenibacillus strains were grown in ISP2 liquid media (10 g/L maltextract, 4 g/L Bacto yeast extract, 4 g/L glucose monohydrate) for 6days at 22° C. at 150 rpm. OD_(600nm) indicating bacterial growth wasmeasured at different time points.

TABLE 5 Bacterial growth of Paenibacillus strains in liquid ISP2 medium.OD at 600 nm Paenibacillus strain 0 d 3 d 6 d Lu17007 0.011 3.110 3.050BD-62 0.013 0.442 0.446

Example 4—In-Vitro Confrontation Assay for Antifungal Activity

Antagonistic activity of the Paenibacillus strains against plantpathogens was shown in in-vitro confrontation assay. The phytopathogenicfungi used are Sclerotina scierotiorum (SCLSCL), Botrytis cinerea(BOTRCI), Alternaria sp. (ALTESP) and Phytophthora infestans (PHYTIN).

As growth medium for BOTRCI, ALTESP, SCLSCL, ISP2 medium is usedcomprising per litre: 10 g malt extract (Sigma Aldrich, 70167); 4 gBacto yeast extract (Becton Dickinson, 212750); 4 g glucose monohydrate(Sigma Aldrich, 16301); 20 g Agar (Becton Dickinson, 214510), pH about7, Aq. bidest. As growth medium for PHYTIN, V8 medium is used comprisingper litre: 200 ml of vegetable juice, 3 g calcium carbonate (MerckMillipore, 1020660250); 30 g Agar (Becton Dickinson, 214510), pH 6.8,Aq. bidest.

The Paenibacillus strains were point-inoculated on one side of an agarplate. An agar block (approx. 0.3 cm²) containing one actively growingplant pathogen was put in the center of the plate. After incubating for7-14 days at 25° C., the growth of the plant pathogen was examined,especially for inhibition zones.

Thereafter, the agar plates are incubated at ° C. for about 7-14 daysbefore evaluation. Antibiosis is scored by evaluation of the diameter ofthe fungi-free zone (zone of inhibition). Competition is scored bycomparing the diameter of the growth of the fungal pathogen on plateswith bacterial strains in comparison to control plates. Mycoparasitismcan be documented in case the bacteria overgrows the fungal pathogen andalso parasitize the pathogens. This can be visualized by microscopy.

The Paenibacillus strains showed antifungal activity against all testedplant pathogens.

TABLE 6 In-vitro confrontation assay results. Diameter of zone ofinhibition [mm] Paenibacillus strain PHYTIN BOTRCI ALTESP SCLSCL Lu167748 2 2 2 Lu17007 8 8 5 2 Lu17015 8 5 5 2 BD-62 2 5 0 0

Example 5—Glasshouse Tests for Activity Against Plant Pathogenic FungiUse Example 5.1: Activity Against Late Blight on Tomato Caused byPhytophthora Infestans with Protective Application

Commercially available young tomato seedlings (“Goldene Königin”) wereused for the described greenhouse trial. 2 replications (pots with 1plant each) were used per treatment. Plants were grown in commerciallyavailable substrate (Universal, Floragard) at approx. 22° C. in thegreenhouse. The humidity was controlled using a special device (˜90%humidity). The plants were sprayed to runoff with crude/whole culturebroth of 6 days old cultures of the respective Paenibacillus strain(depending on the setup) using a spray cabinet. Culture conditions forthe strains are described in Example 3. One day after application thetreated plants were inoculated with a suspension of sporangia ofPhytophthora infestans (PHYTIN). After inoculation, the trial plantswere immediately transferred to a humid chamber. The extent of fungalattack on the leaves was visually assessed 5-7 days after inoculation.Fungal attack in the untreated control was between 80-100% and set to100% for comparison reason.

TABLE 7 Paenibacillus strain PHYTIN (% fungal attack) Lu17007 4 Lu1677420 BD-62 53

Use Example 5.2: Activity Against Grey Mold on Pepper Caused by BotrytisCinerea with Protective Application

Commercially available young pepper seedlings (“Neusiedler Ideal”) wereused for the described greenhouse trial. 2 replications (pots with 1plant each) were used per treatment. Plants were grown in commerciallyavailable substrate (Universal, Floragard) at approx. 22° C. in thegreenhouse. The humidity was controlled using a special device (˜90%humidity). The plants were sprayed to runoff with crude culture broth of6 days old cultures of the respective Paenibacillus strain (depending onthe setup) using a spray cabinet. Culture conditions for the strains aredescribed in Example 3. One day after application the treated plantswere inoculated with a suspension of spores of Botrytis cinerea(BOTRCI). After inoculation, the trial plants were immediatelytransferred to a humid chamber. The extent of fungal attack on theleaves was visually assessed 5-7 days after inoculation. Fungal attackin the untreated control was between 80-100% and set to 100% forcomparison reason.

TABLE 8 Paenibacillus strain BOTRCI (% fungal attack) Lu17007 2 Lu1677416 Lu17015 20 BD-62 97

Use Example 5.3: Activity Against Early Blight on Tomato Caused byAlternaria Solani with Protective Application

Commercially available young tomato seedlings (“Goldene Königin”) wereused for the described greenhouse trial. 2 replications (pots with 1plant each) were used per treatment. Plants were grown in commerciallyavailable substrate (Universal, Floragard) at approx. 22° C. in thegreenhouse. The humidity was controlled using a special device (˜90%humidity). The plants were sprayed to runoff with crude/whole culturebroth of 6 days old cultures of the respective Paenibacillus strain(depending on the setup) using a spray cabinet. Culture conditions forthe strains are described in Example 3. One day after application thetreated plants were inoculated with a suspension of spores of Alternariasolani (ALTESO). After inoculation, the trial plants were immediatelytransferred to a humid chamber. The extent of fungal attack on theleaves was visually assessed 5-7 days after inoculation. Fungal attackin the untreated control was between 80-100% and set to 100% forcomparison reason.

TABLE 9 Paenibacillus strain ALTESO (% fungal attack) Lu17007 3 Lu1701516 BD-62 96

Use Example 5.4: Activity Against Soybean Rust on Soybean Caused byPhakopsora Pachyrhizi with Protective Application

Commercially available young soybean seedlings (“Mentor”) were used forthe described greenhouse trial. 2 replications (pots with 1 plant each)were used per treatment. Plants were grown in commercially availablesubstrate (Universal, Floragard) at approx. 22° C. in the greenhouse.The humidity was controlled using a special device (˜90% humidity). Theplants were sprayed to runoff with crude culture broth of 2-6 days oldcultures of Paenibacillus spp. (depending on the setup) using a spraycabinet. One day after application the treated plants were inoculatedwith a suspension of spores of Phakopsora pachyrhizi (PHAKPA). Afterinoculation, the trial plants were immediately transferred to a humidchamber. The extent of fungal attack on the leaves was visually assessed5-7 days after inoculation.

Use Example 5.5: Activity Against Fusarium Head Blight on Wheat Causedby Fusarium Graminearum with Protective Application

Commercially available young wheat seedlings were used for the describedgreenhouse trial. 2 replications (pots with 1 plant each) were used pertreatment. Plants were grown in commercially available substrate(Universal, Floragard) at approx. 22° C. in the greenhouse. The humiditywas controlled using a special device (˜90% humidity). The plants weresprayed to runoff with crude culture broth of 2-6 days old cultures ofPaenibacillus spp. (depending on the setup) using a spray cabinet.Culture conditions are described in Example 3. One day after applicationthe treated plants were inoculated with a suspension of spores ofFusarium graminearum (GIBBZE). After inoculation, the trial plants wereimmediately transferred to a humid chamber. The extent of fungal attackon the leaves was visually assessed 5-7 days after inoculation.

Use Example 5.6: Activity Against Speckled Leaf Blotch on Wheat Causedby Septoria Tritici with Protective Application

Commercially available young wheat seedlings were used for the describedgreenhouse trial. 2 replications (pots with 1 plant each) were used pertreatment. Plants were grown in commercially available substrate(Universal, Floragard) at approx. 22° C. in the greenhouse. The humiditywas controlled using a special device (˜90% humidity). The plants weresprayed to runoff with crude culture broth of 2-6 days old cultures ofPaenibacillus spp. (depending on the setup) using a spray cabinet.Culture conditions are described in Example 3. One day after applicationthe treated plants were inoculated with a suspension of spores ofSeptoria tritici (SEPTTR). After inoculation, the trial plants wereimmediately transferred to a humid chamber. The extent of fungal attackon the leaves was visually assessed 21-28 days after inoculation.

Use Example 5.7: Activity of the Paenibacillus Cells and of theSupernatant Against Various Pathogens with Protective Application

Whole culture broth from 6 days old cultures of Paenibacillus strainLu17007 was obtained according to Use Example 3 and used as in theexperimental setup of Use Example 5.1 to 5.3. Alternatively, such wholeculture broth was filtered through a filter with 0.2 μm pore size toobtain the culture medium and the crude cell fraction. The crude cellfraction could further be washed three times with the original volumesof phosphate-buffered saline to obtain washed cells.

The glasshouse trials were performed as described in the Use Examples5.1, 5.2 and 5.3 above for the respective pathogens Phytophthorainfestans, Botrytis cinerea and Alternaria solani. The extent of fungalattack on the leaves was visually assessed 5-7 days after inoculation.Fungal attack in the untreated control was between 80-100% and set to100% for comparison reason.

TABLE 10 Paenibacillus % fungal attack by culture component BOTRCIALTESO PHYTIN Whole culture broth 0 2 7 Culture medium 3 40 3 Crude cellfraction 0 5 4 Washed cells 1 10 1

Example 6—Enzymatic Tests Use Example 6.1: Chitinase

Chitinase test solid medium:

2 g/l NaNO₃, 1 g/l K₂HPO₄, 0.5 g/l MgSO₄, 0.5 g/l KCl, 0.2 g/l pepton,15 g/l agar, 10 g/l chitin from crab shells (Sigma-Aldrich C7170).

Test solid medium is autoclaved and filled into 9 cm Petri dishes.Paenibacillus strains are inoculated in the center of the plates andincubated for two days at 27° C. Thereafter, the plates are stained witha 1:3 diluted Lugol solution (Carl Roth N052.2) for 5 to 10 min. Lugolsolution is poured out and the plates are photographed and evaluated.Growth of the different strains was no more than 5-10 mm. Non-stainedzones (correlating with chitinase activity) varied from 0 mm (noactivity; “−” in Table 11) to several cm (“+” in Table 11).

Use Example 6.2: Cellulase

Cellulase test solid medium:

2 g/l NaNO₃, 1 g/l K₂HPO₄, 0.5 g/l MgSO₄, 0.5 g/l KCl, 0.2 g/l pepton,15 g/l agar, carboxymethyl cellulose, sodium salt (Sigma-Aldrich419273).

Medium is autoclaved poured into 9 cm Petri dishes. Paenibacillusstrains are inoculated in the center of the plates and incubated for twodays at 27° C. After incubation plates are stained with a 1:3 dilutedLugol solution (Carl Roth N052.2) for 5 to 10 min. Lugol solution ispoured out and plates photographed.

Use Example 6.3: Amylase

Amylase test solid medium:

2 g/l NaNO₃, 1 g/l K₂HPO₄, 0.5 g/l MgSO₄, 0.5 g/l KCl, 0.2 g/l pepton,15 g/l agar, 10 g/l soluble starch (Merck 1.01252).

Medium is autoclaved poured into 9 cm Petri dishes. Paenibacillusstrains are inoculated in the center of the plates and incubated for twodays at 27° C. After incubation plates are stained with a 1:3 dilutedLugol solution (Carl Roth N052.2) for 5 to 10 min. Lugol solution ispoured out and plates photographed.

TABLE 11 Chitinase, cellulose and amylase activities of Paenibacillusstrains. Strain Chitinase Cellulase Amylase Lu16774 + + − Lu17007 ++ + +Lu17015 + + + BD-62 − − − −, no activity; +, low activity; +, regularactivity; ++, high activity.

Example 7—Fusaricidin-Type Metabolites Obtained from PaenibacillusStrains Example 7.1: Large Scale Cultivation of Bacterial Isolates andExtraction of Fusaricidin-Type Metabolites

a) Cultivation

The Paenibacillus strains were cultivated on agar plates containing GYMmedium (10 g/l glucose, 4 g/l yeast extract, 10 g/l malt extract; pH5.5, adjusted before autoclaving) and 20 g/l agar. Cultivation wasperformed for 10 to 20 days at room temperature. For maintenance agarslants with the same medium were used and stored at 4° C.

Small scale liquid cultures (250 ml GYM medium in 500 ml flasks) wereinoculated with 4-5 pieces of a well grown agar culture and cultivatedin an orbital shaker at 120 rpm at room temperature (20-23° C.).

Large scale fermentations were performed in 20 l fermenters with 15 lGYM medium (total capacity of fermenters was not used because of foamformation) inoculated with 250 ml well grown liquid culture andfermentation was carried out at room temperature (20-23° C.) withagitation (120 rpm) and aeration (3 l/min) for 5 to 8 days.

b) Extraction

One equal volume of isopropanol was added to the whole culture broth (noseparation of biomass from liquid culture was performed). Afteragitation and incubation for 2 to 16 hours, common table salt (sodiumchloride—100 to 200 g/l) was added to the mixture until phase separationof the organic and aqueous phase was visible.

The isopropanol phase was concentrated in vacuo. The resulting extract,still containing large amount of salt, was dissolved in methanol,centrifuged for better precipitation of salt residues, and the organicphase was concentrated again. This step was repeated until no saltprecipitate was present anymore.

c) Purification

i) Silica Gel Chromatography

-   30 grams of extract were dissolved in methanol and bound to 50 g    silica gel (Merck, K60, 70-230 mesh), dried at 40° C. and layered    onto 1 kg of silica gel (column 10 cm diameter, 30 cm high approx.).-   Elution was carried out in four steps as following:    Step 1—4 l ethyl acetate    Step 2—4 l ethyl acetate:methanol (3:1, v/v)    Step 3—7 l ethyl acetate:methanol (1:1, v/v)    Step 4—4 l methanol-   The third fraction (intermediate 1), containing the active    compounds, was dried in vacuo and dissolved in 40% methanol (MeOH)    in 0.1% formic acid (FA) (concentration: 100 mg/ml). The other    fractions were discarded.

ii) Chromabond HR-X Fractionation

-   20 ml of intermediate 1 was loaded onto a previously equilibrated    (with 40% MeOH in 0.1% FA) Chromabond HR-X cartridge    (Macherey-Nagel, 1000 mg, ref 730941). The cartridge was washed with    100 ml 40% MeOH in 0.1% FA and eluted with 60 ml 70% MeOH in 0.1%    FA. This intermediate 1-1 was then dried in vacuo.

iii) Preparative HPLC on a Sunfire C18 Column

-   Intermediate 1-1 was dissolved in DMSO (concentration: 200 mg/ml)    and 300 μl of intermediate 1-1 were chromatographed on a Sunfire C18    column (19×250 mm, 5 μm, Waters) as follows:    16 min at 10 ml/min, isocratic 70% 0.2 FA; 30% acetonitrile (ACN),    1 min at 14 ml/min, gradient to 65% 0.2% FA; 35% ACN,    5 min at 14 ml/min, isocratic 65% 0.2% FA; 35% ACN.

Five fractions could be detected. All five resulting fractions weredried in vacuo and dissolved in DMSO (concentration: 125 mg/ml). Furtherpurification was performed using the same column and isocraticconditions (flow: 10.5 ml/min) adjusted for every fraction (12.5 mg perrun):

-   -   Fraction 1:69% 0.2 FA; 31% ACN; two peaks detected (1-1 and 1-2)    -   Fraction 2: 69% 0.2 FA; 31% ACN; two peaks detected (2-1 and        2-2)    -   Fraction 3: 69% 0.2 FA; 31% ACN; three peaks detected (3-1, 3-2        and 3-3)    -   Fraction 4/5: 67% 0.2 FA; 33% ACN; one peak detected (4/5)    -   Fraction 6: 65% 0.2 FA; 35% ACN; two peaks detected (6-1 and        6-2)

The purity and quantity of the following samples was sufficient for NMRanalysis and structure elucidation: peaks 1-2, 2-1, 3-2, 4/5 and 6-1.

Example 7.2: Structural Elucidation of Compounds 1A and 1 B

From peak 2-1 of fraction 2, a mixture of compounds 1A and 1 B (ratioabout 3:7) was obtained as a brown oil ([α]_(D) ²⁵=+20.9 (c=0.6,DMSO-d₆)).

The molecular formula C₄₇H₇₈N₁₀O₁₂ of the major component, compound 1B,was deduced from the HR-ESI-MS spectrum which gave a peak at m/z975.5863 [M+H]⁺; ESI-MS: 975.6 (100%, [M+H]⁺), 488.4 (51%, [M+2H]²⁺).

Besides, the mixture also contained as minor component, the lighterhomologue 1A, and the mass difference between both compounds was 14 amu.This observation was supported by a second peak observed in the ESI-MSspectrum at m/z 961.6.

The NMR spectra (Table 12) included in addition to signals ofexchangeable protons between δ6.83 and 8.58, resonances of carbonyl inthe range of δ166.0-174.5 and methine signals between δ47.8 and δ60.4indicative for a peptide.

Extensive analysis of the 1 D- and 2D-NMR data of compound 1 B revealedthe presence of six amino acids including tyrosine (Tyr), glutamine(Gln), alanine (Ala), two threonines (Thr1 and Thr2) and isoleucine(Ile). Their sequence was found using two or three bonds correlationsacross amide functions. Thus, COSY, NOESY (FIG. 2) and HMBC (FIG. 3)spectra depicted correlations from the nitrogen-proton of Thr2 at δ8.58to the signal of methine proton of Thr2 at δ3.84 and the carbonyl atδ166.7 of Tyr while the same relationship was noted between thenitrogen-proton of Tyr at δ8.52 and the signal of methylene proton ofTyr at δ2.60 and the carbonyl at δ170.4 of Ile. Furthermore, the methinehydrogen of Ile at δ4.16 had a strong correlation with the carbonylsignal of Ile at δ170.4 and a weak contact with that of Thr1 at δ168.6;the signal of thep-nnethine proton at δ5.30 of Thr1 correlated with thecarbonyl signal at δ170.4 of Ala. Additionally to the aforementionedcorrelations, others were displayed from the N-proton at δ7.27 of Ala tothe methine proton at δ4.20 of the same amino acid while this latterproton had the same interaction with the carbonyl of its amino and theone of Gln. Besides, a cross peak was revealed from the exchangeableproton at δ8.20 of Gln to the methine hydrogen at δ3.87 of Gln and thecarbonyl of Thr2 at δ170.6; these above-mentioned data suggested thecyclodepsipeptidic structure for compound 1B.

This cyclodepsipeptide 1B contained a terminal guanidine β-hydroxy fattyacid attached to Thr1 since a key correlation was observed between thesignal of its α-methine proton at δ4.39 and the resonance of a carbonylat δ171.9; HMBC contacts from that carbonyl at δ171.9 to the α-methyleneprotons at δ2.35 and thep-nnethine proton at δ3.77 were further observedas well as between the methylene protons at δ3.03 and the guanidinecarbon at δ157.2. The side chain was deduced to contain twelve methylenegroups between the β-hydroxy and the guanidine group on the basis of thefragment ion observed in the APCI-MS-MS spectrum of the parent [M+H]⁺ion at m/z 256.2. Likewise, this spectrum provided information (FIG. 4b) which confirmed the connection sequence of amino acids and led toelucidate the structure of compound 1B as shown in FIG. 1.

Signals of a CH₂ group at 2.80, 2.52/36.3 in the 1D- and 2D-spectracorresponded presumably to the β-CH₂ group of asparagine (Asn) incompound 1A. This conclusion was supported by reported data(Heterocycles 53, 1533-1549, 2000) in conjunction to fragments obtainedfrom MS/MS of the parent peak at m/z 961.6 (FIG. 4a ). Likewise, thelatter analyses provided information (FIGS. 4a, 4b ) which confirmed theconnection sequence of amino acids in both compounds and led toelucidate the structure of compounds 1A and 1B as shown in FIG. 1.

Example 7.3: Structural Identification of Compounds 2A and 2B asFusaricidins C and D

From peak 1-2 of fraction 1, a mixture of compounds 2A and 2B (ratioabout 1:1) was obtained as a brown oil. The molecular formula of theheavier component, compound 2B, was determined to be C₄₆H₇₆N₁₀O₁₂ on thebasis of the low resolution mass spectrometry. Analysis of the NMR data(Table 13) allowed to identify compound 2B as fusaricidin D. The lightercomponent of the mixture, compound 2A, was likewise identified asfusaricidin C, in which the Gln residue of fusaricidin C is replaced byAsn.

The mass spectrometric fragmentation pattern of the parent ions of m/z961.6 and 947.6 for compounds 2B and 2A, respectively, (FIGS. 5a, 5b )confirmed the length of the substituted fatty acid side chain to beidentical as in compound 1B. Fusaricidins C and D have formerly beenreported by Kajimura et al. (J. Antibiot. 50, 220-228, 1997).

Example 7.4: Structural Identification of Compound 3 as LI-F08b

From peak 6-1 of fraction 6, compound 3 was isolated as a brown oil andits low resolution presented a peak at m/z 925.6 [M+H]⁺ which, combinedwith NMR data (Table 14), led to the molecular formula C₄₄H₈₀N₁₀O₁₁.Compound 3 showed similar features in the NMR spectra as compound 1B andcompound 2B (fusaricidin D) except for the presence of aromatic signals(Table 14). Thus, characteristic resonances of a peptide were observednamely ten signals of protons attached to nitrogen between δ 6.89 and8.49, eight resonances of carbonyl ranged between δ168.1 and 174.3, andsix signals of N-methine comprised between δ48.0 and 59.5. A detailedanalysis of the HMQC, COSY and TOCSY spectra revealed the presence ofsix amino acids including Gln, two units of Thr, two units of Ile andAla. Furthermore, these spectra showed chemical shifts attributable tothe same β-hydroxyl fatty acid with a terminal guanidine as in compounds1A, 1B and fusaricidins C (2A) and D (2B). The position of this sidechain was determined on the basis of a long range correlation found onthe HMBC spectrum between the proton signal of N-methine at δ4.44 ofThr1 and the carbonyl signal at δ172.1 of the fatty acid. The sequenceof the amino acids was deduced from NOESY interactions and thefragmentation pattern (FIG. 6).

The combination of the NMR data (Table 14) and mass spectrometry led toidentify the metabolite compound 3 as LI-F08b, herein also calledfusaricidin LI-F08b, reported for the first time by Kuroda et al.(Heterocycles 53, 1533-1549, 2000).

Example 7.5: Structural Identification of Compounds 4A and 4B as LI-F06aand LI-F06b and of Compounds 5A and 5B as Rusaricidin A and B,Respectively

From peak 4/5 of fraction ⅘, a mixture of two further metabolites,compounds 4A and 4B (ratio about 1:3), was obtained which gave two peaksat m/z 897.5 (4A) and 911.6 (4B) in the ESI-MS spectrum, suggesting twofurther homologous cyclodepsipeptides. Resonances indicative forpeptides were observed in their NMR spectra (Table 15) as well as thoseof a β-hydroxyl fatty acid terminating in a guanidine group. Thefragmentation patterns of both parent ions found for compounds 4A and 4B(FIGS. 7a, 7b ) allowed to determine the sequence of amino acids and toidentify the constituents of the mixture as LI-F06a (4A) and LI-F06b(4B), respectively.

Obtained from peak 3-2 of fraction 3, the mixture of compounds 5A and 5B(ratio about 1:3) was analyzed in the same manner. The ESI mass spectrumof the mixture showed two peaks at m/z 883.6 (5A) and 897.5 (5B) and thefragmentation patterns of these parent ions (FIGS. 8a, 8b ) inconjunction to NMR data (Table 16) allowed to identify the components asfusaricidin A (5A) and fusaricidin B (56). The data found for 4A, 4B, 5Aand 5B matched those previously reported. (J. Antibiot. 50, 220-228,1997; Heterocycles 53, 1533-1549, 2000).

TABLE 12 ¹H (DMSO-d₆, 600 MHz) and ¹³C-NMR (DMSO-d₆, 150 MHz) data ofcompounds 1A and 1B. Compounds 1 *Pos. δ_(H) δ_(c) Compound 1A Thr1 NH7.79 (br) — 1 — 168.6 2 4.46 (br d, 8.5) 56.4 3 5.30 (overlapped) 70.2 41.13 (overlapped) 16.6 Ala NH 7.22 (br) — 1 — nf* 2 4.13 (overlapped)47.7 3 1.11 (overlapped) 17.8 Asn NH 8.33 (overlapped) — 1 — 169.7 24.20 (1H, m) 50.6 3 2.52 (m), 36.3 2.80 (dd, 5.9, 15.1) 4 — 172.5 5 — —NH₂ 6.99 (br s), 7.42 (br s) — Thr2 NH 8.50 (overlapped) — 1 — 170.6 23.94 (m) 59.9 3 3.94 (m) 65.5 4 1.05 (br) 20.3 Tyr NH 8.48 (overlapped)— 1 — nf 2 4.60 (m) 54.2 3 2.60 (overlapped) 36.8 2.88 (overlapped) 4 —127.7 5 and 9 7.07 (d, 8.7) 130.2 6 and 8 6.60 (overlapped) 114.7 7 —155.9 Ile NH 7.28 (br s) — 1 — nf 2 4.16 (overlapped) 56.5 3 1.34(overlapped) 37.2 4 1.34 (overlapped) 25.4 5 0.52 (overlapped) 14.4 60.59 (overlapped) 11.4 *FA 1 — 171.9 2 2.35 (overlapped) 43.1 3 3.77(overlapped) 67.5 4 1.34 (overlapped) 36.8 5-12 1.19-1.30 (br s)29.0-29.2 13  1.25 (br s) 21.2 14  1.43 (overlapped) 28.7 15  3.03(overlapped) 40.6 *Gu NH nf — 16  — 157.2 Compound 1B Thr1 NH 8.18 (brs) — 1 — 168.6 2 4.39 (br d, 8.7) 56.9 3 5.30 (m) 70.2 4 1.13 (d, 6.4)16.7 Ala NH 7.27 (br s) — 1 — 170.4 2 4.20 (m) 47.8 3 1.17 (d, 7.1) 17.8Gln NH 8.20 (br s) — 1 — 170.4 2 3.87 (m) 53.2 3 1.96 (m), 2.08 (m) 26.24 2.08 (m), 2.18 (m) 32.0 — — 174.3 NH₂ 6.83 (br s), 7.26 (br s) — Thr2NH 8.58 (br s) — 1 — 170.6 2 3.84 (m) 60.5 3 3.85 (m) 65.8 4 1.08(overlapped) 20.0 Tyr NH 8.52 (br s) — 1 — 166.7 2 4.51 (m) 54.5 3 2.60(m), 2.88 (m) 36.9 4 — 127.8 5 and 9 7.06 (d, 8.5) 130.2 6 and 8 6.60(d, 8.5) 114.7 7 — 155.9 Ile NH 7.42 (br s) — 1 — 170.4 2 4.16 (br d,8.5) 56.5 3 1.34 (m) 37.2 4 1.22 (m), 1.34 (m) 25.4 5 0.53 (overlapped)14.4 6 0.61 (overlapped) 11.4 FA 1 — 171.9 2 2.35 (m) 43.3 3 3.77 (m)67.5 4 1.34 (m) 36.9 5-12 1.19-1.30 (br s) 29.0-29.2 13  1.25 (br s)21.2 14  1.43 (m) 28.5 15  3.03 (q, 6.6) 40.6 Gu NH 8.40 (br s) — 16  —157.2 *Pos. = position; FA = fatty acid; Gu = Guanidine; nf = not found.Legend applies also to Tables 13 to 16.

TABLE 13 ¹H (DMSO-d₆, 600 MHz) and ¹³C-NMR (DMSO-d₆, 150 MHz) data ofcompounds 2A and 2B. Compounds 2 = fusaricidins C and D Pos. δ_(H) δ_(C)Compound 2A = fusaricidin C Thr1 NH 7.66 (d, 7.1) — 1 — 168.5 2 4.44 (brd, 8.9) 56.6 3 5.31 (m) 70.2 4 1.13 (overlapped) 16.5 Ala NH 7.21 (br) —1 — nf 2 4.12 (m) 47.7 3 1.12 (overlapped) 17.8 Asn NH 8.26 (br) — 1 —169.7 2 4.21 (m) 50.5 3 2.53 (overlapped), 36.3 2.80 (dd, 6.3, 15.0) 4 —172.6 5 — — NH₂ nf — Thr2 NH 8.52 (overlapped) — 1 — 170.3 2 3.85 (m)60.5 3 3.86 (m) 65.8 4 1.09 (d, 5.7) 19.9 OH-3 4.96 (br d, 4.2) — Tyr NH8.46 (overlapped) — 1 — nf 2 4.60 (m) 54.2 3 2.63 (overlapped) 36.9 2.87(overlapped) 4 — 127.7 5 and 9 7.08 (overlapped) 130.2 6 and 8 6.60(overlapped) 114.7 7 — 155.8 OH nf — Val NH 7.30 (overlapped) — 1 — nf 24.12 (br s) 57.5 3 1.59 (m) 30.9 4 0.56 (d, 6.4) 18.2 5 0.35 (d, 6.5)18.7 FA 1 — nf 2 2.37 (overlapped) 43.1 3 3.79 (overlapped) 67.5 4 1.35(overlapped) 36.9 5 1.22 (overlapped) 25.3 6-12 1.20-1.27 (br s)29.1-29.2 13  1.26 (br s) 26.1 14  1.44 (overlapped) 28.5 15  3.07(overlapped) 40.7 Gu NH nf — 16  — 156.8 Compound 2B = fusaricidin DThr1 NH 8.17 (br s) — 1 — 168.6 2 4.40 (br d, 8.9) 57.0 3 5.30 (m) 70.34 1.14 (overlapped) 16.7 Ala NH 7.60 (br s) 1 — 170.6 2 4.19 (m) 47.8 31.17 (d, 7.2) 17.7 Gln NH 8.08 (br s) — 1 — 170.4 2 3.86(m) 53.2 3 1.98(m), 26.1 2.09 (m) 4 2.10 (m), 31.9 2.18 (m) 5 — 174.3 NH₂ 6.84 (br s),— 7.28 (br s) Thr2 NH 8.47 (overlapped) — 1 — 170.6 2 3.94 (m) 59.9 33.92 (m) 65.7 4 1.05 (d, 5.8) 20.2 OH-3 5.05 (d, 2.9) — Tyr NH 8.52(overlapped) — 1 — 172.3 2 4.52 (m) 54.6 3 2.63 (m), 36.9 2.87 (m) 4 —127.7 5 and 9 7.06 (d, 8.4) 130.2 6 and 8 6.60 (d, 8.4) 114.7 7 — 155.8OH 9.13 (br s) — Val NH 7.42 (br s) — 1 — 170.3 2 4.12 (br s) 57.5 31.59 (m) 31.0 4 0.57 (d, 6.3) 18.3 5 0.40 (d, 6.6) 18.7 FA 1 — 172.0 22.37 (m) 43.3 3 3.79 (m) 67.6 4 1.35 (m) 36.9 5 1.22 (br s) 25.3 6-121.20-1.27 (br s) 29.1-29.2 13  1.26 (br s) 26.1 14  1.44 (m) 28.7 15 3.07 (q, 6.7) 40.7 Gu NH 7.60 (br s) — 16  — 156.8

TABLE 14 ¹H (DMSO-d₆, 600 MHz) and ¹³C-NMR (DMSO-d₆, 150 MHz) data ofcompound 3 being LI-F08b. Compound 3 = LI-F08b Pos. δ_(H) δ_(C) Thr1 NH7.55 (br s) — 1 — 168.1 2 4.44 (br d, 8.4) 56.6 3 5.33 (m) 70.2 4 1.15(d, 6.5) 16.7 Ala NH 7.53 (br s) — 1 — 170.6 2 4.05 (m) 48.0 3 1.22 (brs) 17.2 Gln NH 7.93 (br s) — 1 — 170.5 2 3.94 (m) 52.7 3 1.98 (m), 26.52.09 (m) 4 2.12 (m), 31.9 2.20 (m) 5 — 174.3 NH₂ 6.89 (br s), — 7.32 (brs) Thr2 NH 8.48 (br s) — 1 — 170.7 2 4.03 (m) 59.5 3 3.98 (m) 65.7 41.08 (d, 6.1) 19.8 Ile1 NH 8.49 (br s) — 1 — 172.5 2 4.15 (t, 7.6) 57.33 1.81 (m) 35.4 4 1.17 (m), 24.4 1.41 (m) 5 0.80 (t, 6.3) 10.6 6 0.81(d, 7.2) 15.5 Ile2 NH 7.30 (br s) — 1 — 171.3 2 4.53 (m) 55.3 3 1.65 (m)38.2 4 1.01 (m), 25.5 1.37 (m) 5 0.83 (t, 6.4) 11.4 6 0.70 (d, 7.4) 14.2FA 1 — 172.1 2 2.37 (d, 5.7) 43.4 3 3.77(m) 67.6 4 1.37 (m) 36.9 5-121.20-1.28 (br s) 29.0-29.2 13  1.25 (br s) 26.2 14  1.43 (m) 28.7 15 3.03 (q, 6.7) 40.6 Gu NH 8.37 (br s) — 16  — 157.2

TABLE 15 ¹H (DMSO-d₆, 600 MHz) and ¹³C-NMR (DMSO-d₆, 150 MHz) data ofcompounds 4A and 4B. Compounds 4 = LI-F06a and LI-F06b Pos. δ_(H) δ_(C)Compound 4A = LI-F06a Thr1 NH 8.31 (br) — 1 — 168.5 2 4.40 (m) 56.9 35.30 (m) 70.5 4 1.14 (m) 16.6 Ala NH nf — 1 — 170.6 2 3.97 (m) 47.9 31.15 (overlapped) 17.3 Asn NH 8.06 (br) — 1 — 169.8 2 4.28 (m) 50.5 32.55 (m), 36.9 2.75 (dd, 6.7, 15.1) 4 — 172.6 5 — — NH₂ nf — Thr2 NH8.54 (br) 1 — 170.4 2 3.91 (m) 60.5 3 3.92 (m) 65.6 4 1.09 (d, 6.4) 19.6Val NH 7.28 (m) — 1 — nf 2 4.40 (overlapped) 57.3 3 1.83 (overlapped)32.0 4 0.75 (d, 6.6) 18.1 5 0.84 (overlapped) 19.3 Ile NH 7.31(overlapped) — 1 — nf 2 4.51 (overlapped) 55.5 3 1.65 (overlapped) 38.14 1.02 (m), 25.4 1.36 (m) 5 0.82 (overlapped) 15.6 6 0.72 (overlapped)14.4 FA 1 — 172.1 2 2.44 (dd) 43.1 3 3.81 (m) 67.7 4 1.37 (overlapped)36.9 5-12 1.22-1.24 (br s) 29.1-29.2 13  1.25 (br s) 26.4 14  1.43 (m)28.5 15  3.03 (q, 6.7) 40.7 Gu NH nf — 16  — 157.2 Compound 4B = LI-F06bThr1 NH 7.59 (br s) — 1 — 168.4 2 4.44 (m) 56.7 3 5.32 (m) 70.3 4 1.15(m) 16.6 Ala NH 7.53 (br s) 1 — 170.7 2 4.07 (m) 48.0 3 1.21 (d, 7.3)17.4 Gln NH 7.96 (br s) — 1 — 170.7 2 3.93 (m) 52.9 3 1.97 (m), 26.52.10 (m) 4 2.12 (m), 32.0 2.21 (m) 5 — 174.4 NH₂ 6.88 (br s), — 7.33 (brs) Thr2 NH 8.48 (br) — 1 — 170.6 2 4.02 (m) 59.7 3 3.99 (m) 65.7 4 1.08(d, 6.4) 19.8 Val NH 7.39 (m) — 1 — 171.0 2 4.39 (m) 57.0 3 1.83 (m)31.6 4 0.74 (d, 6.6) 18.4 5 0.80 (overlapped) 19.2 Ile NH 7.23(overlapped) — 1 — 171.2 2 4.51 (1H, m) 55.6 3 1.65 (m) 38.1 4 1.02 (m),25.5 1.36 (m) 5 0.82 (overlapped) 15.6 6 0.71 (overlapped) 14.3 FA 1 —172.2 2 2.37 (m) 43.4 3 3.78 (m) 67.7 4 1.37 (m) 36.9 5 1.22-1.24 (br s)29.1-29.2 13 1.25 (br s) 26.2 14 1.43 (m) 28.5 15 3.03 (q, 6.7) 40.7 GuNH 8.34 (br s) — 16 — 157.2

TABLE 16 ¹H (DMSO-d₆, 600 MHz) and ¹³C-NMR (DMSO-d₆, 150 MHz) data ofcompounds 5A and 5B. Compounds 5 = fusaricidins A and B, LI-F04a andLI-F04b Pos. δ_(H) δ_(C) Compound 5A = fusaricidin A Thr1 NH 7.66 (br) —1 — 168.5 2 4.46 (m) 56.6 3 5.32 (m) 70.4 4 1.16 (overlapped) 16.3 AlaNH 7.26 (br) — 1 — 170.6 2 4.00 (m) 47.8 3 1.15 (overlapped) 17.4 Asn NH8.10 (br) — 1 — 169.8 2 4.28 50.5 3 2.53 (m), (q, 6.6) 36.7 2.76 (dd,6.6, 15.0) 4 — 172.5 5 — — NH₂ nf — Thr2 NH 8.54 (br s) — 1 — nf 2 3.91(m) 60.4 3 3.91 (m) 65.6 4 1.09 (d, 5.6) 19.6 Val NH nf — 1 — nf 2 4.40(m) 57.1 3 1.82 (m) 31.4 4 nf nf 5 0.82 (d, 6.0) 19.1 Val NH 8.41 (br s)— 1 — 172.1 2 4.13 (m) 58.3 3 2.02 (m) 29.7 4 0.86 (d, 6.7) 18.2 5 0.84(7.0) 19.3 FA 1 — 172.1 2 2.37 (br d, 5.8) 43.4 3 3.80 (m) 67.6 4 1.37(m) 36.9 5-12 1.22-1.25 (br s) 26.2-29.2 13  1.25 (br s) 26.2 14  1.43(m) 28.7 15  3.03 (q, 6.7) 40.6 Gu NH nf — 16  — 157.2 Compound 5B =fusaricidin B Thr1 NH 8.30 (d, 8.0) — 1 — 168.4 2 4.40 (br d, 8.5) 57.03 5.31 (m) 70.3 4 1.15 (d, 5.7) 16.6 Ala NH 7.53 (br s) — 1 — 170.6 24.10 (m) 47.9 3 1.20 (d, 7.2) 17.5 Gln NH 8.53 (d, 4.3) — 1 — 170.6 23.92 (m) 52.9 3 1.98 (m), 26.4 2.09 (m) 4 2.10 (m), 31.9 2.20 (m) 5 —174.3 NH₂ 6.86 (br s), — 7.30 (br s) Thr2 NH 8.46 (d, 6.9) — 1 — 170.5 24.0 (m) 59.7 3 3.99 (m) 65.5 4 1.07 (d, 6.0) 19.9 Val NH 7.29 (br s) — 1— 171.2 2 4.40 (m) 57.1 3 1.82 (m) 31.5 4 0.76 (d, 6.6) 18.4 5 0.81 (d,6.2) 19.1 Val NH 8.37 (d, 7.6) — 1 — 173.1 2 4.23 (m) 57.8 3 1.99 (m)30.2 4 0.86 (d, 6.7) 18.2 5 0.84 (7.0) 19.3 FA 1 — 172.0 2 2.34 (dd,7.0, 13.5), 43.4 2.44 (dd, 4.9, 13.5) 3 3.80 (m) 67.6 4 1.37 (m) 36.85-12 1.22-1.25 (br s) 26.2-29.2 13  1.25 (br s) 26.2 14  1.43 (m) 28.515  3.03 (q, 6.7) 40.6 Gu NH 8.47 (br s) — 16  — 157.2

No hydrolysis experiments were carried out to determine theconfiguration of the constituting amino acids.

Example 8—Metabolites Produced by Paenibacillus Strains Example 8.1:Production of Metabolites by Paenibacillus Strains

The presence of fusaricidins in general and in particular of thefusaricidins A, B, C, D, LI-F06a, LI-F06b, LI-F08b, 1A and 1 B wasdetermined for the Paenibacillus strains following the procedural stepswhich are described in Example 7.1 above.

TABLE 17 Fusaricidin-type metabolite production of the Paenibacillusstrains. Compound/Fusaricidin 1A 1B 2A 2B 3 4A 4B 5A 5B Strains 1A 1B CD Ll-F08b Ll-F06a Ll-F06b A B Lu16774 + ++ ++ ++ ++ − − ++ ++ Lu17007 +++ ++ ++ ++ + ++ ++ ++ Lu17015 ++ ++ ++ ++ ++ ++ ++ ++ ++ BD-62 − − − −− − − − − Legend: −, compound not detectable; +, compound detectable;++, compound detectable at higher amounts compared to scale +.

The whole culture broth of all of the Paenibacillus strains Lu16774,Lu17007 and Lu17015 contained at least one fusaricidin identified inExample 7 (Table 17). None of these fusaricidins were detected in thewhole culture broth of P. peonae strain BD-62.

The whole culture broth of the Paenibacillus strains Lu16774, Lu17007and Lu17015 all contained the fusaricidins 1A and 1 B. Further, thewhole culture broth of the Paenibacillus strains Lu16774, Lu17007 andLu17015 all contained the fusaricidins A, B, C and D as well as LI-F08b.In addition, the whole culture broth of the Paenibacillus strainsLu17007 and Lu17015 contained fusaricidins LI-F06a and LI-F06b.

Fusaricidins 1A and 1 B were not detected in the whole culture broth ofthe closely related P. peonae strain BD-62. Fusaricidins A, B, C and D,LI-F06a, LI-F06b and LI-F08b were also not in the whole culture broth ofP. peonae strain BD-62.

Example 9: Activity of Metabolites by Paenibacillus Strains AgainstVarious Fungal Pathogens

The fusaricidins A, B, D, 1A and 1 B were obtained were used in thefollowing experiments.

Fungal growth assays were performed in 96 well plates with sporesuspension of the pathogen Botrytis cinerea (BOTRCI, in YBA [10 g Bactopeptone (Becton Dickinson 211677), 10 g yeast extract (Becton Dickinson212750), 20 g sodium acetate, ad 1000 mL aqua bidest] or Alternariasolani (ALTESO, in YBG [10 g Bacto peptone (Becton Dickinson 211677), 10g yeast extract (Becton Dickinson 212750), 20 g glycerine 99%, ad 1000mL aqua bidest]). Fusaricidins and compounds 1A and 1B were dissolvedand diluted in DMSO. Different concentrations ranging from 60 μM down to0.3 μM were pipetted into the microtiter plate. An aqueous suspension of10⁴ spores/ml was added. The plates were incubated at about 18° C.Fungal growth was determined by measuring the optical density at 600 nmin a microplate reader 3 and 7 days after the inoculation of the sporesand compared to the untreated control (DMSO). IC₅₀ (concentration [μM]of the respective metabolite required for 50% inhibition of fungalgrowth) has been determined thereafter.

Notably, the compounds 1A and 1B showed the highest antifungal efficacywith IC₅₀ values of 0.4-0.6 μM (Tab. 18).

TABLE 18 Antifungal growth inhibition of Paenibacillus metabolites IC₅₀values Compound/Fusaricidin 1A 1B 2B 5A 5B Pathogen 1A 1B Fus. D Fus. AFus. B (Evaluation day) Fungal growth inhibition (IC50 [μM]) ALTESO (3d) 0.6 0.6 1.1 1.3 1.1 ALTESO (7 d) 0.5 0.4 0.6 0.7 0.6 BOTRCI (7 d) 0.30.4 0.5 0.5 0.6

In addition, glasshouse trials were performed with fusaricidins 1A and1B as described in the Use Examples 5.1 to 5.5 above for the respectivepathogens Botrytis cinerea (BOTRCI), Alternaria solani (ALTES0),Phytophthora infestans (PHYTIN), Phakopsora pachyrhizi (PHAKPA) andFusarium graminearum (GIBBZE). The extent of fungal attack on the leaveswas visually assessed 5-7 days after inoculation.

Notably, compounds 1A and 1B were effective in controlling importantfungal diseases on crop plants already at dose levels as low as 7.2 ppmand showed higher antifungal efficacy than Fusaricidin A, B and D(Tables 19 to 21).

TABLE 19 Antifungal efficacy of metabolites determined in planta. %efficacy (% fungal attack) Metabolite tested Conc. BOTRCI ALTESO PHYTINPHAKPA GIBBZE Untreated — 0 (100) 0 (100) 0 (100) 0 (100) 0 (100)Compound 1A 360 ppm 99 100 95 49 Compound 1A  36 ppm 97 74 Compound 1B360 ppm 100  100 97 Compound 1B  36 ppm 97

TABLE 20 Efficacy of metabolites against late blight on tomato caused byPhytophthora infestans with protective application. % efficacyMetabolite tested Conc. (% fungal attack) Untreated — 0 (100)Fusaricidin A 7.2 ppm 15 Fusaricidin B 7.2 ppm 4 Fusaricidin D 7.2 ppm 0Compound 1B 7.2 ppm 44

TABLE 21 Efficacy of metabolites against head blight on wheat caused byFusarium graminearum with protective application. % efficacy Metabolitetested Conc. (% fungal attack) Untreated — 0 (100) Fusaricidin A 360 ppm31 Fusaricidin B 360 ppm 0 Compound 1A 360 ppm 49

TABLE 22 Efficacy of metabolites against head blight on wheat caused bySeptoria tritici with protective application. % efficacy Metabolitetested Conc. (% fungal attack) Untreated — 0 (100) Fusaricidin D 360 ppm50 Compound 1B 360 ppm 80

Example 10: Comparison of Activity of Paenibacillus Polymyxa nov. ssp.Plantarum Strains Lu16674 and Lu17007 with Paenibacillus Polymyxa nov.ssp. Plantarum M-1 Against Various Pathogens in Glass House Trials

Whole culture broth from 6 days old cultures of Paenibacillus strainLu17007, Lu16674 and M1 was obtained according to Use Example 3 and usedas in the experimental setup of Use Example 5.1 to 5.5. The glasshousetrials were performed as described in the Use Examples 5.1 to 5.5 abovefor the respective pathogens. The extent of fungal attack on the leaveswas visually assessed 5-7 days after inoculation.

Notably, the Paenibacillus strains Lu16774 and Lu17007 were effective incontrolling important fungal diseases on crop plants even at highdilution factors and showed higher antifungal efficacy than the closelyrelated strain M-1 (Tables 22 to 27).

TABLE 22 Dilution factor of BOTRCI % efficacy Paenibacillus strain wholeculture broth (% fungal attack) Untreated 0 (100) Lu16674 1:10 95 M-11:10 86 Lu16674 1:50 76 Lu17007 1:50 98 M-1 1:50 51

TABLE 23 Dilution factor of BOTRCI % efficacy Paenibacillus strain wholeculture broth (% fungal attack) Untreated 0 (100) Lu17007 undiluted 92M-1 undiluted 87 Lu17007 1:10 84 M-1 1:10 53 Lu17007 1:50 63 M-1 1:50 32

TABLE 24 Dilution factor of ALTESO % efficacy Paenibacillus strain wholeculture broth (% fungal attack) Untreated 0 (100) Lu16674 1:10 77 M-11:10 41

TABLE 25 Dilution factor of PHYTIN % efficacy Paenibacillus strain wholeculture broth (% fungal attack) Untreated 0 (100) Lu17007 1:10 83 M-11:10 42 Lu16674 1:50 13 Lu17007 1:50 30 M-1 1:50  0

TABLE 26 Dilution factor of PHAKPA % efficacy Paenibacillus strain wholeculture broth (% fungal attack) Untreated 0 (100) Lu17007 undiluted 94M-1 Undiluted 87

TABLE 27 Dilution factor of GIBBZE % efficacy Paenibacillus strain wholeculture broth (% fungal attack) Untreated 0 (100) Lu17007 undiluted 70M-1 undiluted 31 Lu16674 1:50 52 Lu17007 1:50 33 M-1 1:50 24

Examples Concerning the Mixtures and Compositions of the InventionExample 11—Activity Against Early Blight Caused by Alternaria Solani(Alteso) in Microtests

The active compounds were formulated separately as a stock solutionhaving a concentration of 10000 ppm in dimethyl sulfoxide. PaenibacillusLU17007 was used as an exprerimental formulation and diluted with waterto the stated concentration of the active compound. The product Bacillusamyloliquefacies MBI 600 was used as commercial finished formulationsand diluted with water to the stated concentration of the activecompound.

The stock solutions were mixed according to the ratio, pipetted onto amicro titer plate (MTP) and diluted with water to the statedconcentrations. A spore suspension of Alternaria solani in an aqueousbiomalt or yeast-bactopeptone-glycerine solution was then added. Theplates were placed in a water vapor-saturated chamber at a temperatureof 18° C. Using an absorption photometer, the MTPs were measured at 405nm 7 days after the inoculation.

The measured parameters were compared to the growth of the activecompound-free control variant (100%) and the fungus-free and activecompound-free blank value to determine the rela-relative growth in % ofthe pathogens in the respective active compounds. These percentages wereconverted into efficacies.

Calculation of the Expected Efficacy (E_(Colby)) Using the Colby'sFormula

The expected efficacies of active compound combinations were determinedusing Colby's formula (Colby, S. R. “Calculating synergistic andantagonistic responses of herbicide combinations”, Weeds 15, pp. 20-22,1967) and compared with the observed efficacies.E _(Colby) =P _(A) +P _(B) −P _(A) *P _(B)/100  Colby's Formula

E_(Colby) expected efficacy, expressed in % of the untreated control,when using the mixture of the active compounds A and B at theconcentrations a and b

P_(A) efficacy, expressed in % of the untreated control, when using theactive compound A at the concentration a

P_(B) efficacy, expressed in % of the untreated control, when using theactive compound B at the concentration b.

Calculation of the Synergy Factor (SF)

For a determination of synergism the Synergy Factor (SF) between theobserved experimental efficacy of the mixtures E_(measured) and theexpected efficacy of the mixture E_(Colby) is calculated asSF=E _(measured) /E _(Colby)

A Synergy Factor greater or smaller than 1 indicates a deviation fromthe hypothesis of independent action which means that biologically thetwo components act together or against each other. If SF>1, synergism isobserved; if SF<1, antagonism is observed.

Alteso Calculated Active Concen- efficacy Syner- compound/ trationObserved according to gism SF active mixture (ppm/cfu) efficacy Colby(%) (%) factor Fusaricidin 1b 0.063 13 MBI 600 1.6 × 10³ cfu 17Fusaricidin 1b 0.063 47 28 19 1.68 MBI 600 1.6 × 10³ cfu

The documents as cited herein are incorporated by reference.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Compounds 1A, 1B, 2A, 2B, 3, 4A, 4B, 5A and 5B.

FIG. 2. Key NOESY and COSY correlations of compound 1B.

FIG. 3. HMBC correlation of compound 1B.

FIG. 4. Fragmentation patterns a) of compound 1A and b) of compound 1B.

FIG. 5. Fragmentation patterns a) of compound 2A (fusaricidin C) and b)of compound 2B (fusaricidin D).

FIG. 6. Fragmentation pattern of compound 3 (LI-F08b).

FIG. 7. Fragmentation patterns a) of compound 4A (LI-F06a) and b) ofcompound 4B (LI-F06b).

FIG. 8. Fragmentation patterns a) of compound 5A (fusaricidin A) and b)of compound 5B (fusaricidin B).

FIG. 9 shows the percentage identity of the complete 16S rDNA sequenceof the Paenibacillus strains of the invention to related taxa aftermultiple sequence alignment. Legend: * Strain numbers: 1=Paenibacillusstrain Lu16774; 2=Paenibacillus strain Lu17015; 3=Paenibacillus strainLu17007; 4=Paenibacillus peoriae NRRL BD-62; 5=Paenibacillus americanusMH21; 6=Paenibacillus brasiliensis PB172; 7=Paenibacillus campinasensis324; 8=Paenibacillus chibensis JCM 9905; 9=Paenibacillus glucanolyticusDSM 5162; 10=Paenibacillus hunanensis FeL05; 11=Paenibacillus jamilaeCECT 5266; 12=Paenibacillus kribbensis AM49; 13=Paenibacillus lactis MB1871; 14=Paenibacillus lautus JCM 9073; 15=Paenibacillus macerans IAM12467; 16=Paenibacillus massiliensis 2301065; 17=Paenibacillus pabuliHSCC 492; 18=Paenibacillus peonae DSM 8320 (BD-57); 19=Paenibacilluspini 522; 20=Paenibacillus polymyxa IAM 13419; 21=Paenibacilluspurispatii ES_MS17; 22=Paenibacillus sediminis GT-H3; 23=Paenibacillusterrae AM141; 24=Paenibacillus terrigena A35; 25=Paenibacillustimonensis 2301032; 26=Paenibacillus turicensis MOL722; 27=Paenibacillusuliginis N3/975; 28=Cohnella thermotolerans CCUG 47242. Strains 6 to 28are type strains for the respective species.

Similarities of the strains Lu16774, Lu17007 and Lu17015 withPaenibacillus peoriae (NRRL BD-62 and DSM 8320) have been marked in boldletters.

FIG. 10 shows a phylogenetic dendrogram calculated from the % identityof 16S-rDNA sequences of the Paenibacillus strains of the invention withother taxa (FIG. 9). The root of the tree was determined by includingthe 16S rRNA gene sequence of Cohnella thermotolerans into the analysis.The scale bar below the dendrogram indicates 1 nucleotide substitutionsper 100 nucleotides.

FIG. 11 shows the RiboPrint pattern obtained from samples of thePaenibacillus strains of the invention in comparison to a sample of theclosely related P. peoriae strain BD-62 using Ribo-RiboPrinter MicrobialCharacterization System and a phylogenetic dendrogram resultingtherefrom.

FIG. 12 shows the percentage identity of the DNA sequence of the dnaNgene of the Paenibacillus strains of the invention to relatedPaenibacillus strains after multiple sequence alignment.

Legend: * Strain numbers: 1=Paenibacillus strain Lu16774;2=Paenibacillus strain Lu17007; 3=Paenibacillus strain Lu17015; 4=P.peoriae DSM 8320T=KCTC 3763^(T) (GenBank acc. no. AGFX00000000; J.Bacteriol. 194, 1237-1238, 2012); 5=P. polymyxa 1-43 (GenBank acc. no.ASRZ01000000; deposition no. GCMCC 4965; CN 102352332 B); 6=P. polymyxaA18 (GenBank acc. no JWJJ00000000.1; NCBI Project ID 225496); 7=P.polymyxa ATCC 842^(T)=DSM 36^(T)=KCTC 3858T (GenBank acc. no.AFOX00000000; J. Bacteriol. 193(18), 5026-5027, 2011); 8=P. polymyxaCF05 (GenBank acc. no. CP009909; Genome Announc 3(2):e00198-15.Doi:10.1128/genomeA.00198-15); 9=P. polymyxa CICC 10580 (GenBank acc.no. JNCB00000000; Genome Announc. 2(4):e00854-14.doi:10.1128/genomeA.00854-14); 10=P. polymyxa DSM 365 (GenBank acc. no.JMIQ00000000; J. Biotechnol. 195, 72-73, 2015); 11=P. polymyxa E681(GenBank acc. no. CP000154; GenomeNet Ref Seq NC_014483.2; J. Bacteriol.192(22), 6103-6104, 2010); 12=P. polymyxa M-1 (GenBank acc. no.HE577054.1; GenomeNet Ref Seq NC_017542.1); 13=P. polymyxa NRRL B-30509(GenBank acc. no. JTHO00000000; Genome Announc. 2015 March-April; 3(2):e00372-15); 14=P. polymyxa SC2 (GenBank acc. no. CP002213; J. Bacteriol.193 (1), 311-312, 2011); 15=P. polymyxa SQR-21 (GenBank acc. no.CP006872; GenomeNet Ref Seq NZ_CP006872.1; Genome Announc. 2014March-April; 2(2): e00281-14); 16=P. polymyxa Sb3-1 (GenBank acc. no.CP010268; Genome Announc. 2015 March-April; 3(2): e00052-15); 17=P.polymyxa TD94 (GenBank acc. no. ASSA00000000); 17=P. polymyxa WLY78(GenBank acc. no. ALJV00000000); P. terrae HPL-003 (GenBank acc. no.CP003107; NCBI Ref Seq NC_016641.1); P. polymyxa CR1 (GenBank acc. no.CP006941; Genome Announc. 2014 January-February; 2(1): e01218-13).

FIG. 13 shows the percentage identity of the DNA sequence of thecomplete gyrB gene of the Paenibacillus strains of the invention torelated Paenibacillus strains after multiple sequence alignment. Strainnumbers are described in Legend to FIG. 12.

FIG. 14 shows the percentage identity of the DNA sequence of thecomplete recF gene of the Paenibacillus strains of the invention torelated Paenibacillus strains after multiple sequence alignment. Strainnumbers are described in Legend to FIG. 12.

FIG. 15 shows the percentage identity of the DNA sequence of thecomplete recN gene of the Paenibacillus strains of the invention torelated Paenibacillus strains after multiple sequence alignment. Strainnumbers are described in Legend to FIG. 12.

FIG. 16 shows the percentage identity of the DNA sequence of thecomplete rpoA gene of the Paenibacillus strains of the invention torelated Paenibacillus strains after multiple sequence alignment. Strainnumbers are described in Legend to FIG. 12.

FIG. 17 shows the maximum likelihood denrogram on basis of the completednaN gene sequence of strains of the P. polymyxa complex. The scale of0.1 shown corresponds to 1% nucleotide exchanges.

FIG. 18 shows the maximum likelihood denrogram on basis of the completegyrB gene sequence of strains of the P. polymyxa complex. The scale of0.1 shown corresponds to 1% nucleotide exchanges.

FIG. 19 shows the maximum likelihood denrogram on basis of the completerecF gene sequence of strains of the P. polymyxa complex. The scale of0.1 shown corresponds to 1% nucleotide exchanges.

FIG. 20 shows the maximum likelihood denrogram on basis of the completerecN gene sequence of strains of the P. polymyxa complex. The scale of0.1 shown corresponds to 1% nucleotide exchanges.

FIG. 21 shows the maximum likelihood denrogram on basis of the completerpoA gene sequence of strains of the P. polymyxa complex. The scale of0.1 shown corresponds to 1% nucleotide exchanges.

FIG. 22 shows the Amino Acid Index (AAI) matrix of representativegenomes of the P. polymyxa complex performed according to Example 2.5.Strain numbers are described in Legend to FIG. 12.

The invention claimed is:
 1. A mixture, comprising as activecomponents 1) at least one Paenibacillus strain or a culture mediumthereof, a cell-free extract thereof, or at least one metabolitethereof; wherein the at least one Paenibacillus strain is selected fromthe group consisting of: a) Paenibacillus strain Lu16774 deposited withDSMZ under Accession No. DSM 26969; b) Paenibacillus strain Lu17007deposited with DSMZ under Accession No. DSM 26970; c) Paenibacillusstrain Lu17015 deposited with DSMZ under Accession No. DSM 26971; and d)a Paenibacillus strain which comprises a DNA sequence exhibiting d1) atleast 99.6% nucleotide sequence identity to the DNA sequences SEQ IDNO:4 or SEQ ID NO:9; or d2) at least 99.8% nucleotide sequence identityto the DNA sequence SEQ ID NO:14; or d3) at least 99.9% nucleotidesequence identity to the DNA sequences SEQ ID NO:5 or SEQ ID NO:10; ord4) at least 99.2% nucleotide sequence identity to the DNA sequence SEQID NO:15; or d5) at least 99.2% nucleotide sequence identity to the DNAsequences SEQ ID NO:6 or SEQ ID NO:11; or d6) at least 99.8% nucleotidesequence identity to the DNA sequence SEQ ID NO:16; or d7) at least99.8% nucleotide sequence identity to the DNA sequences SEQ ID NO:7 orSEQ ID NO:12; or d8) at least 99.3% nucleotide sequence identity to theDNA sequence SEQ ID NO:17; or d9) 100.0% nucleotide sequence identity tothe DNA sequences SEQ ID NO:8 or SEQ ID NO:13; or d10) at least 99.9%nucleotide sequence identity to the DNA sequence SEQ ID NO:18; and 2) atleast one biopesticide II selected from the groups L1) to L5): L1) atleast one microbial pesticide with fungicidal, bactericidal, viricidaland/or plant defense activator activity selected from: Ampelomycesquisqualis, Aspergillus flavus, Aureobasidium pullulans, Bacillusaltitudinis, B. amyloliquefaciens, B. megaterium, B. mojavensis, B.mycoides, B. pumilus, B. simplex, B. solisalsi, B. subtilis, B. subtilisvar. amyloliquefaciens, Candida oleophila, C. saitoana, Clavibactermichiganensis (bacteriophages), Coniothyrium minitans, Cryphonectriaparasitica, Cryptococcus albidus, Dilophosphora alopecuri, Fusariumoxysporum, Clonostachys rosea f. catenulate (also named Gliocladiumcatenulatum), Gliocladium roseum, Lysobacter antibioticus, L.enzymogenes, Metschnikowia fructicola, Microdochium dimerum,Microsphaeropsis ochracea, Muscodor albus, Paenibacillus alvei,Paenibacillus polymyxa, Pantoea vagans, Penicillium bilaiae, Phlebiopsisgigantea, Pseudomonas sp., Pseudomonas chloraphis, Pseudozymaflocculosa, Pichia anomala, Pythium oligandrum, Sphaerodesmycoparasitica, Streptomyces griseoviridis, S. lydicus, S.violaceusniger, Talaromyces flavus, Trichoderma asperelloides, T.asperellum, T. atroviride, T. fertile, T. gamsii, T. harmatum, T.harzianum, T. polysporum, T. stromaticum, T. virens, T. viride, Typhulaphacorrhiza, Ulocladium oudemansii, Verticillium dahlia, and/or zucchiniyellow mosaic virus (avirulent strain); L2) at least one biochemicalpesticide with fungicidal, bactericidal, viricidal and/or plant defenseactivator activity selected from: harpin protein and/or Reynoutriasachalinensis extract; L3) at least one microbial pesticide withinsecticidal, acaricidal, molluscidal and/or nematicidal activityselected from: Agrobacterium radiobacter, Bacillus cereus, B. firmus, B.thuringiensis, B. thuringiensis ssp. aizawai, B. t. ssp. israelensis, B.t. ssp. galleriae, B. t. ssp. kurstaki, B. t. ssp. tenebrionis,Beauveria bassiana, B. brongniartii, Burkholderia spp., Chromobacteriumsubtsugae, Cydia pomonella granulovirus (CpGV), Cryptophlebia leucotretagranulovirus (CrleGV), Flavobacterium spp., Helicoverpa armigeranucleopolyhedrovirus (HearNPV), Helicoverpa zea nucleopolyhedrovirus(HzNPV), Helicoverpa zea single capsid nucleopolyhedrovirus (HzSNPV),Heterorhabditis bacteriophora, Isaria fumosorosea, Lecanicilliumlongisporum, L. muscarium, Metarhizium anisopliae, Metarhiziumanisopliae var. anisopliae, M. anisopliae var. acridum, Nomuraea rileyi,Paecilomyces fumosoroseus, P. lilacinus, Paenibacillus popilliae,Pasteuria spp., P. nishizawae, P. penetrans, P. ramosa, P. thornea, P.usgae, Pseudomonas fluorescens, Spodoptera littoralisnucleopolyhedrovirus (SpliNPV), Steinernema carpocapsae, S. feltiae, S.kraussei, Streptomyces galbus, and/or S. microflavus; L4) at least onebiochemical pesticide with insecticidal, acaricidal, molluscidal,pheromone and/or nematicidal activity selected from: L-carvone, citral,(E,Z)-7,9-dodecadien-1-yl acetate, ethyl formate, (E,Z)-2,4-ethyldecadienoate (pear ester), (Z,Z,E)-7,11,13-hexadecatrienal, heptylbutyrate, isopropyl myristate, lavanulyl senecioate, cis-jasmone,2-methyl 1-butanol, methyl eugenol, cis-jasmone, methyl jasmonate,(E,Z)-2,13-octadecadien-1-ol, (E,Z)-2,13-octadecadien-1-ol acetate,(E,Z)-3,13-octadecadien-1-ol, R-1-octen-3-ol, pentatermanone,(E,Z,Z)-3,8,11-tetradecatrienyl acetate, (Z,E)-9,12-tetradecadien-1-ylacetate, Z-7-tetradecen-2-one, Z-9-tetradecen-1-yl acetate,Z-11-tetradecenal, Z-11-tetradecen-1-01, extract of Chenopodiumambrosiodes, Neem oil, and/or Quillay extract; L5) at least onemicrobial pesticide with plant stress reducing, plant growth regulator,plant growth promoting and/or yield enhancing activity selected from:Azospirillum amazonense, A. brasilense, A. lipoferum, A. irakense, A.halopraeferens, Bradyrhizobium spp., B. elkanii, B. japonicum, B.liaoningense, B. lupini, Delftia acidovorans, Glomus intraradices,Mesorhizobium spp., Rhizobium leguminosarum by. phaseoli, R. I. bv.trifolii, R. I. bv. viciae, R. tropici, and/or Sinorhizobium meliloti,wherein the mixture comprises component 1) and component 2) in asynergistically effective amount.
 2. The mixture of claim 1, whereinsaid at least one Paenibacillus strain has antifungal activity againstat least two of the plant pathogens selected from the group consistingof Alternaria spp., Botrytis cinerea, Phytophthora infestans andSclerotinia sclerotiorum.
 3. The mixture of claim 1, wherein said atleast one Paenibacillus strain is capable of producing at least one ofthe following compounds:

in a growth medium comprising at least one source of carbon and onesource of nitrogen.
 4. The mixture of claim 1, wherein the component 1)comprises a substantially purified culture of said at leastonePaenibacillus strain as defined in claim
 1. 5. The mixture of claim 1,wherein the component 1) comprises a whole culture broth, a culturemedium or a cell-free extract of said at least comprises a whole culturebroth, a culture medium or a cell-free extract of at least onePaenibacillus strain as defined in claim
 1. 6. The mixture of claim 1,wherein the component 1) comprises said at least one metabolite of theat least one Paenibacillus strain selected from the group consisting ofpolymyxins, octapeptins, polypeptins, pelgipeptins and fusaricidins. 7.The mixture of claim 6, wherein the at least one metabolite is selectedfrom fusaricidins of formula I

wherein R is selected from the group consisting of15-guanidino-3-hydroxypentadecanoic acid (GHPD) and12-guanidinododecanoic acid (12-GDA); X¹ is threonine; X² is selectedfrom the group consisting of isoleucine and valine; X³ is selected fromthe group consisting of tyrosine, valine, isoleucine and phenylalanine;X⁴ is threonine; X⁵ is selected from the group consisting of glutamineand asparagine; X⁶ is alanine; and wherein an arrow defines a single(amide) bond either between the carbonyl moiety of R and the amino groupof the amino acid X¹ or between the carbonyl group of one amino acid andthe amino group of a neighboring amino acid, wherein the tip of thearrow indicates the attachment to the amino group of said amino acid X¹or of said neighboring amino acid; and wherein the single line withoutan arrow head defines a single (ester) bond between the carbonyl groupof X⁶ and the hydroxyl group of X¹.
 8. The mixture of claim 7, whereinthe at least one metabolite is selected from fusaricidins of formula I

wherein R is selected from the group consisting of15-guanidino-3-hydroxypentadecanoic acid (GHPD) and12-guanidinododecanoic acid (12-GDA); X¹ is threonine; X² is isoleucine;X³ is tyrosine; X⁴ is threonine; X⁵ is selected from the groupconsisting of glutamine and asparagine; X⁶ is alanine; and wherein anarrow defines a single (amide) bond either between the carbonyl moietyof R and the amino group of the amino acid X¹ or between the carbonylgroup of one amino acid and the amino group of a neighboring amino acid,wherein the tip of the arrow indicates the attachment to the amino groupof said amino acid X¹ or of said neighboring amino acid; and wherein thesingle line without an arrow head defines a single (ester) bond betweenthe carbonyl group of X⁶ and the hydroxyl group of X¹.
 9. The mixture ofclaim 8, wherein the at least one metabolite is selected fromfusaricidins 1A and 1 B:


10. A composition comprising the mixture of claim 1 and an auxiliary.11. The composition of claim 10, further comprising a pesticide IIIselected from the groups SF) and SI): SF) Fungicides inhibitors ofcomplex III at Q_(o) site selected from the group consisting of:pyraclostrobin, azoxystrobin, picoxystrobin, trifloxystrobin,dimoxystrobin, enestroburin, fenaminstrobin, fluoxastrobin,kresoxim-methyl, mandestrobine, metominostrobin, orysastrobin,pyrametostrobin, and pyraoxystrobin; broad-spectrum pyridine andpyrazole inhibitors of complex II selected from the group consisting of:fluxapyroxad, boscalid, benzovindiflupyr, penflufen, penthiopyrad,sedaxane, fluopyram, bixafen, and isopyrazam; Basidiomycetes-specificinhibitors of complex II selected from the group consisting of:carboxin, benodanil, fenfuram, flutolanil, furametpyr, mepronil,oxycarboxin, and thifluzamide; ATP production inhibitor silthiofam;fungicidal azole compounds selected from the group consisting of:ipconazole, difenoconazole, prothioconazole, prochloraz, triticonazole,flutriafol, cyproconazole, diniconazole, diniconazole-M,fluquinconazole, flusilazole, hexaconazole, imazalil, imibenconazole,metconazole, myclobutanil, simeconazole, tebuconazole, triadimenol,uniconazole, thiabendazole; Oomycetes fungicides selected from:oxathiapiprolin, valifenalate, metalaxyl, metalaxyl-M, ethaboxam,dimethomorph, zoxamide, flumorph, mandipropamid, pyrimorph,benthiavalicarb, and iprovalicarb; MAP/histidine kinase inhibitor:fludioxonil; benzimidazole compounds selected from the group consistingof: thiophanate-methyl, and carbendazim; dithiocarbamate compoundsselected from the group consisting of: thiram, and ziram; SI)Insecticides GABA antagonist compounds selected from the groupconsisting of: fipronil, ethiprole, vaniliprole, pyrafluprole,pyriprole, and5-amino-1-(2,6-dichloro-4-methyl-phenyl)-4-sulfinamoyl-1H-pyrazole-3-carbothioicacid amide; lepidopteran-specific ryanodine receptor inhibitors selectedfrom the group consisting of: chlorantraniliprole and flubendiamide;cross-spectrum ryanodine receptor inhibitor: cyantraniliprole;pyrethroid sodium channel modulators selected from the group consistingof: tefluthrin, bifenthrin, cypermethrin, alpha-cypermethrin,cyfluthrin, beta-cyfluthrin, lambda-cyhalothrin, deltamethrin,esfenvalerate, etofenprox, fenvalerate, flucythrinate, and permethrin;systemically-active neonicotinoid compounds: clothianidin, imidacloprid,thiamethoxam, dinotefuran, acetamiprid, flupyradifurone, thiacloprid,triflumezopyrim, nitenpyram; Acetylcholinesterase inhibitors, chloridechannel activators and sulfoximines: sulfoxaflor, acephate,chlorpyrifos, thiodicarb, abamectin, spinosad; other insecticide:tioxazafen.
 12. A plant propagation material having a coating comprisingthe composition of claim
 10. 13. A method of controlling, suppressingpathogens or preventing pathogen infection, wherein the pathogens, theirhabitat or the materials or plants to be protected against pathogenattack, or the soil or propagation material are treated with aneffective amount of the mixture of claim
 1. 14. A method of controlling,suppressing pathogens or preventing pathogen infection, wherein thepathogens, their habitat or the materials or plants to be protectedagainst pathogen attack, or the soil or propagation material are treatedwith an effective amount of the composition of claim 10.